Catalogue English

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PRICE QUALITY and SERVICES
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S U M M A R Y
PAGE
ALPHABETIE INDEX BIOCHEMICAL ELECTROLYTES BIOCHEMICAL ENZYMES BIOCHEMICAL SUBSTRATES HEMOSTASIS / HEMATOLOGY IMMUNOHEMATOLOGY / HEMATOLOGY SEROLOGY AND QUICK - TESTS 8 - 13 16 - 23 26 - 39 42 - 47 50 - 54 56 - 63
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ALPHABETIC INDEX
DESIGNATION ALAT. GPT ALBUMINE ALKALIN PHOSPHATASE ALP (DGKC) AMYLASE AMINO TRANSFERASE ANTI A MONOCLONAL ANTIBODY ANTI AB - MONOCLONAL ANTIBODY ANTI B - MONOCLONAL ANTIBODY APTT ASAT - GOT ASO LATEX AUTO FIBRI 1 AUTO FIBRI 2 CALCIUM CALIBRATOR FOR TOTAL AND DIRECT BILIRUBIN Rf 20046 20094 20015 20031 20041 30111 30131 30121 30021 20042 40037 30033 30034 20051 201010 Page 16 26 17 19 18 50 51 52 42 20 56 43 44 8 27
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DESIGNATION CHOLESTEROL CHOLESTEROL-HDL CREATININE CREATINE KINASE CRP LATEX DIRECT BILIRUBIN FIBRINOGEN GAMMA GT GLUCOSE HEMOGLOBIN IRON FERROZINE LDH MAGNESIUM PHOSPHORUS COLORIMETRIC PHOSPHORUS UV METHOD POLYVALENTE ANTIGLOBULIN RHEUMATOID FACTORS LATEX STREPTOLYSIN TPHA TRIGLYCERIDES Rf 20111 20113 20151 20006 40034 20102 30031 20021 20121 30041 20061 20011 20071 20081 20083 30181 40036 40021 40011 20138 Page 28 29 30 21 57 31 46 22 32 47 9 23 10 11 12 54 58 59 61 36
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DESIGNATION Rf Page
TOTAL AND DIRECT BILIRUBIN TOTAL BILIRUBIN TOTAL PROTEINS TOTAL IRON BINDING CAPACITY THROMBOPLASTIN UCG 25 UREA (BERTHELOT) UREA (KINETIC METHOD) URIC ACID
20101 20103 20161 20062 30011 40032 20141 20142 20091
33 34 35 13 45 62 37 38 39
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BIOCHEMICAL ELECTROLYTES
Page
CALCIUM
Rf. 20051 Rf. 20054 2 x 160 ml 2 x 125 ml
IRON FERROZINE
Rf. 20061 Rf. 20064 2 x 100 ml 5 x 100 ml
MAGNESIUM
Rf. 20071 Rf. 20074 INORGANIC PHOSPHORUS : COLORIMETRIC METHODS Rf. 20081 Rf. 20082 Rf. 20084 100 tests 200 tests 250 tests 2 x 160 ml 2 x 125 ml
10
11
INORGANIC PHOSPHORUS UV METHOD

Rf. 20083 320 tests
12
TOTAL IRON - BINDING CAPACITY

Rf. 20062 Rf. 20063 50 tests 100 tests
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PRESENTATION
Ref. 20051 (320 Tests) R1 : 2 x 80 ml R2 : 2 x 80 ml R3 : 1 x 4 ml Ref. 20054 (250 Tests) R1 : 1 x 125 ml R2 : 1 x 125 ml R3 : 1 x 3 ml
CALCIUM
Colorimetric Test O- cresolphtaleine
PRINCIPLE
Calcium with O-cresolphtaleine complex compound, at alkaline pH, yields a red colored complex, which intensity is proportional to the calcium concentration.
CALCU LATION
OD. Sample xn OD.Standard mg/dl n= 10 mg/l n = 100 mmol/l n = 2.5
REAGENTS
Reagent 1 2- Amino-2-methyl Buffer solution 1- propanol Reagent 2 Chromogen solution Reagent 3 Standard Gresolphtaleine compl. 8-hydroxyquinoleine 500 mmol/l
LINEARITY
This method is linear up to 15 mg/dl (150 mg/l) If is greater, dilute the sample 1 : 2 with saline solution
0.62 mmol/l 69 mmol/l
and multiply the result by 2.
REFERENCES VALUES
Calcium standard 10 mg/dl 100 mg/l 2,5 mmol/l Serum Newborn Children Adults 7.5-12 mg/dl 1.87-3 mmol/l 10.0-11.0 mg/dl 2.5-2.87 mmol/l 9.0-10.6 mg/dl 2.25-2.65 mmol/l Urines Newborn Children 1.8 mglkg/24 h 0.025-0.2 mmol/kg/24h 2.6 mglkg/24 h 0.05-0.150 mmol/kg/24h 150-300 mg/24 h Adult 3.5-7.5 mmol/24h
PREPARATION AND STABILITY

Mix 1 volume of buffer solution (R1) with 1 volume of chromogen solution (R2) Stability : 4 hours at 20-25C 20 hours at 2-8C
SAMPLES
Serum, heparinized plasma. Urine diluted 1/3 with distilled water acidified to pH 3-4 with dilute HCL.
PROCEDURE
Wavelength : .........................................570 nm (550-590) Temperature : ...........................................................20-25C Cuvette : .........................................................1 cm lightpath
NOTES
Blank Standard Sample Reacton mixture 1 ml Standard 20 l 1 ml Sample 20 l 1 ml As Calcium is a widley diffused ion, essential precaution must be taken against accidental contaminations, possible using disposable materials. Traces of chelating agents such as EDTA, wich may be present in detergents, sometimes prevent the formation of the coloured complex.
Mix and wait 5 min. at room temperature Read the extinction (E) of standard and sample against the blank. The colour is stable for at least 1 hour.
BIBLIOGRAPHY
Ste, J. and Lewis W.H.P. Clin Chim Acta 2, 576 (1957).
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Oct 2006
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PRESENTATION
Ref. 20061 (100 Tests) R1 : 2 x 100 ml R2 : 1 x 2,5g R3 : 1 x 5 ml R4 : 1 x 10 ml Ref. 20064 (250 Tests) R1 : 5 x 100 ml R2 : 1 x 5g R3 : 1 x 12 ml R4 : 2 x 10 ml
IRON
Colorimetric test (FERROZINE)
PRINCIPLE
The Iron dissociated from transferrin-iron complex by a solution of guanidine acetate and reduced by ascorbic acid reacts with ferrozine to give a pink complex.
REAGENTS
Reagent 1 Reagent 2 Reagent 3 Reagent4 Guanidine, HCI Acetate buffer pH 5 Ascorbic acid Ferrozine Standard 4,5 mmol/l
Reagent Standard Sample Sample blank blank Distilled water 200 l Standard R4 200 l Sample 200 l 200 l Reagent A 1 ml Reagent B 1 ml 1 ml 1 ml Mix and let stand for 10 minutes at +20-25C, then read the absorbance. The colour of reaction is stable for at least 30 minutes.
40 mmol/l 1 mg/l 17,9 mol/l
CALCULATION
OD sample - OD sample blank Iron = x n OD standard mg/l : n = 1 mol/l : n = 17,9

Reagent A : Add one measure (250 mg) of reagent 2 at 50 ml of reagent 1. Reagent B : (Chromogen buffer) Mix 1 volume (ex 40 I) of reagent 3 with 25 volumes (ex : 1 ml) of reagent A Stability : 2 Weeks at 2-8C 3 days at 20-25C
LINEARITY
This method is linear up to 1000 g/100 ml (10 mg/l - mol/l).
REFERENCES VALUES
Men Women 69-158 g/dl 12,5-28,3 mol/l 59-145 g/dl 10,7-26 mol/l
SAMPLE
Serum or heparinized plasma free of hemolysis.
Anyway each laboratory should assign their own reference values.
NOTES
* All glassware must be iron-free : wash with diluled hydrocloric acid and rinse with distilled water. Do not use dark glass containers for the conservation of the solution. Use preferably plastic throway tubes. * Do not use hemolysed samples. Use heparine at the suggested quantities as anticoagulant : high leveles can produce torbidness in the reaction mixture. * Copper and other trace elements dont interfere in the reaction. * The suggested reaction volumes can be proportionaly varied with no change in the calculation.
PROCEDURE
Wavelength : Temperature : Cuvette : Measurement : Read 562 nm (530-590) 20-25C 1 cm light path
- Against reagent blank for standard and sample. - Against reagent A for sample blank. the solutions must be brought at +20 to +25c before use. Prepare standard and a reagent blank for each series of determination.
BIBLIOGRAPHY
Persijn et al Chem. Acta 35,91 (1971). Stoockey L. anal. Chem. 42, 779 (1970) Williams et al. Clin. Chem. 23, 237 (1977).
FT An 02
Oct 2006
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PRESENTATION
Ref. 20071 (320 Tests) R1 : 2 x 80 ml R2 : 2 x 80 ml R3 : 1 x 5 ml Ref. 20074 (250 Tests) R1 : 1 x 125 ml R2 : 1 x 125 ml R3 : 1 x 3 ml
MAGNESIUM
Calmagite Method
PRINCIPLE
Magnesium forms a purple colored complex when treated with calmagite in alkaline solution. In presence of EGTA, the reaction is specific. The intensity of the purple colour is proportional to the magnesium concentration.
CALCULATION
O.D. Sample Magnesium : ------------------------- x n O.D. Standard mg/dl mg/l mmol/l Urine : multiply by dilution factor n=2 n = 20 n = 0.824
REAGENTS
Reagent 1 Amino-Methyl propanol EGTA Calmagite Magnesium Standard 1 mmol/l 0.20 mmol/l 0.30 mmol/l 2 mg/dl 20 mg/l 0.824 mmol/l
Reagent 2 Reagent 3
LINEARITY
Up to 5 mg/dl (50 mg/l) (2.06 mmol/l)
REFERENCES VALUES
Serum, plama : 1.6 - 2.5 mg/dl 16 - 25 mg/l 0.65 -1.05 mmol/l Urine : 0.65- 12.5 mmol/24h

Mix 1 volume of buffer solution (R1) with 1 volume of calmagite solution (R2). Stability : 24 hours at 20-25C 4 days at 2-8C
SAMPLES
Serum, heparinized plasma. Urine diluted 1/10 with distilied water, acidified to pH 3-4 with dilute HCI.
NOTES
Use only plastic material
PROCEDURE
Wavelength : Temperature : Cuvette : Read against reagent blank 520 nm (500-550) 20-25C 1 cm light path
BIBLIOGRAPHY
Gindler E., Clin. Chem. 17, 662 (1971).
Blank Standard Sample Working reagen 1 ml
Standard 10 l 1ml
Sample
10 l 1 ml
Mix and read the optical density (O.D.) after incubation for 5 minutes at room temperature Stability of the colour : 1 hour
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Juin 2008
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PRESENTATION
Ref. 20081 (100 Tests) R1 : 1 x 100 ml R2 : 1 x 100 ml R3 : 1 x 4 ml R4 : 1 x 70 ml Ref. 20082 (200 Tests) R1 : 2 x 100 ml R2 : 2 x 100 ml R3 : 1 x 10 ml R4 : 1 x 120 ml Ref. 20084 (250 Tests) R1 : 2 x 125 ml R2 : 2 x 125 ml R3 : 1 x 10 ml R4 : 1 x 125 ml
INORGANIC PHOSPHORUS
Colorimetric methods
Pipette into test tube :
PRINCIPLE Inorganic phosphorus reacts with ammonium molybdate in presence of sulfuric Acid to form a phosphomolybdate complex. The color of molybdenum blue is proportional to the phosphorus concentration.
Reagent blank Desionised water standard R3 Serum or urine 1/5 Working reagent 50 l
Standard
Sample
50 l 2 ml 2 ml 50 l 2 ml
Mix well and wait 2 mn (presence of trouble in sample). REAGENTS R1: Reductant. Hydroxylamin chloride Sulfuric acid PVP Ammonium heptamolybdate Standard Sodium hydroxyde 2 N R4 0.5 ml 0.5 ml 0.5 ml
0,14 mmol/l 89,63 mmol/ 10 g/l 6,07 mmol/l 1.61 mmol/l 50 mg/l
Mix and let stand for 15 mn at room temperature. Read at 680 nm. The coloration is stable for 1 hour.
R2 : R3: R4 :
CALCULATION Conc. of inogarnic phosphorus : O D Sample --------------------x n O D Standard n = 50 mg/l = 5 mg/dl n = 1.61 mmol/l Urine : Multiply result by 5.
WORKING REAGENT-STABILITY Mix together equal volumes of R1 and R2. Stability 4 hours at 20 -25C 1 week at 2 - 8C SAMPLE Serum or heparinized plasma. Urine diluted 1/5 in distilled water. PROCEDURE Wavelength : Cuvette : Temperature : Measurernent :
LINEARITY The methods is linear up to 150 mg/l (4,8 mmol/l). Samples with higher concentrations should be diluted and reassayed. REFERENCE VALUES Serum Adults 1.10 - 2.10 mmol/l 43 - 65 mg/l 0.90 - 1.45 mmol/l 28 - 45 mg/l 12 - 42 mmol/24 h 0,272 - 1,30 g/24 h
Children
Urine 680 nm 1 cm light path 20-25C against reagent blank
NOTE Use only plastic material. BIBLIOGRAPHY Clim. Chim Acta 15, 155, (I967)
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FT An 04
Oct 2006
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PRESENTATION
Ref. 20083 (320 Tests) R1 : 4 x 80ml R2 : 1 x 10ml
INORGANIC PHOSPHORUS
UV METHOD
PRINCIPLE
Inorganic phosphorus reacts with ammonium molibdate in presence of sulfuric Acid to form a phosphomolibdate complex. Ammonium Molibdate + Sulfuric acid phosphorus phosphomolibdate complex. The absorbance of this complex at 340 nm is proportionnal to the phosphorus concentration.
CALCULATION
[Phosphorus] = O D sample O.D. Standart mg/dl: mg/l: mmol/l: Urine: n=5 n=50 n = 1,61 Multiply the result by 10 xn
REAGENTS
Reagent 1 Sulfuric Acid 200 mmol/l 0.40 mmol/l Ammonium Molibdate Detergent 0.S Standard 5 mg/dl 50 mg I I 1.61 mmol/l
LINEARITY
The method is linear up to 200 mg/l (20 mg/dl - 6,46 mmol/l).
Reagent 2
REFERENCE VALUES
Serum Adults 2,5 - 5,0 mg/dl 25 - 50 mg/l 0,81 - 1,61 mmol/l 4,0 - 7,0 mg/l 40 - 70 mg/l 1,29 - 2,26 mmol/l 16,5 - 48,5 mmol/24 h 0,5-1,5 g /24 h
PREPARATION OF REAGENTS AND STABILITY

All reagents are ready to use. Stable until expiry date printed on the label. Children
SAMPLES
Serum or plasma not hemolyzed Urine dilute 1/10 in distilled water Urine
PROCEDURE
Wavelenght Temperature Cuvette Measurement Pipette into test tube Reagent Blank Standard Sample Working Reagent 1 ml Standard Sample 340 nm 20-25c 1 cm light path Against reagent blank
NOTES
If the sample is lipemic, make a blank by mixing 10 l of sample and 1 ml NaCI solution at 9 g/l and read at 340 nm.
BIBLIOGRAPHY
Daly J.A., Ertingshaussen G., Clin. Chem., 18, 263 (1972) NB : The standard is in acquous solution. Preferentially, use calibrator with serum origin (particularly if an automate is used).
10 l 1 ml
10 l 1 ml
Mix, wait 5 mn and read at 340 nm. The coloration is stable for 30 mn.
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Oct 2006
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PRESENTATION
Ref. 20062 (50 Tests) R1 : 2 x 50 ml R2 : 2 x 5g Ref. 20063 (100 Tests) R1 : 2 x 100ml R2 : 2 x 10g
TOTAL IRON-BINDING CAPACITY

Transfer the supernatant and determine the iron concentration with the iron kit, code 20061and 20064. If Iron and Total Iron binding capacity are prevused, prefere starting by the transferrin saturation step than jointly realise Iron and total iron binding capacity tests.
Auxiliary Kit: Ref : 20061 : Iron (Ferrozine) Ref : 20064 : Iron (Ferrozine)
PRINCIPLE
An excess of iron is added to the serum to saturate the transferrin. The unbound iron is precipitated with basic magnesium carbonate. After centrifugation the iron in the supernatant is determined with the IRON Ref : 20061 ou SFBC iron Ref : 20063
CALCULATION
Multiplie the result per 3.
REFERENCE VALUES
Serum 249-412 g/dl 44.5 - 73.5 mol/l
REAGENTS
Reagent 1 Saturating Iron solution Reagent 2 Adsorbant Iron 5 mg / l 89,5 mol/l
Note : Use preferably plastic throwaway material.
Basic magnesium carbonate
PREPARATION OF WORKING SOLUTIONS AND STABILITY

Reagents ready to use. The expiry date of each reagent is printed on the label.
SAMPLE
Serum or heparinized plasma. Specimen can be stored 7 days at + 2-8C or many weeks at - 20C
PROCEDURE
Sample Reagent 1 1 ml 2 ml
Mix and let stand for 5 minutes at least at + 15 to + 25C; Add Reagent 2 2 measures (200 mg)
Mix carefully, turning over to cuvette. Let stand at 15 to 25C for 20 minutes mixing frequently during this period, then centrifuge for 10 minutes at about 3000 rpm.
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Oct 2006
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BIOCHEMICAL ENZYMES Page

ALAT - GPT
Ref. 20046 Ref. 20047 Ref. 20040 20 x 3 ml 10 x10 ml 9 x 50 ml 17 20 x 3 10 x10 4 x 30 5 x 50 ml ml ml ml 18 16
ALKALIN-PHOSPHATASE DGKC
Ref. Ref. Ref. Ref. 20015 20016 20017 20018
AMINO TRANSFERASES COLORIMETRIC METHOD

Ref. Ref. Ref. Ref. 20039 20041 20048 20049 200 Tests 100 Tests 100 Tests 100 Tests
AMYLASE
Ref. 20031 Ref. 20032 Ref. 20033 20 x 3 ml 30 x12 ml 10 x 10 ml
19
ASAT - GOT
Ref. 20042 Ref. 20043 Ref. 20050 20 x 3 ml 10 x 10 ml 9 x 50 ml
20
CREATINE KINASE
Ref. 200063 Ref. 200071 Ref. 200072 GAMMA GT Ref. 20021 Ref. 20022 20 x 3 ml 10 x 10 ml 60 Tests 150 Tests 180 Tests
21
22
LDH
Ref. Ref. Ref. Ref.
15
23 20011 20012 200123 200125 20 x 3 10 x 10 9 x 50 15 x 10 ml ml ml ml
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PRESENTATION
Ref. 20046 (60 Tests) R1 : 1 x 65ml R2 : 20 x 3 ml (Iyoph) Ref. 20C47 (100 Tests) R1 : 1 x 110ml R2 : 10 x 10 ml (lyoph) Ref. 20040 (450 tests) R1 : 9 x 50ml R2 : 9 x 50 ml (Lyoph)
Kinetic Test. IFCC Without Pyridoxal phosphate
ALAT - GPT
PRINCIPLE Kinetic determination of Alanine Amino transferase activity Method recommended by the IFCC. GPT 2-Oxoglutarate + L-Alanine ----------> Glutamate + Pyruvate LDH Pyruvate + NADH + H+ -----------> L-Lactate + NAD+ The rate of NADH consumption is determined photometrically and is directly proportional to the GPT activity in the sample. GPT : Glutamic pyruvate transaminase LDH : Lactate dehydrogenase REAGENTS Reagent 1 Bufferreagent Reagent 2 Substrate Tris buffer pH 7.5 L-Alanine NADH LDH Oxoglutarate 100 mmol/I 500 mmol/l 0.18 mmol/l 1200 U/l 15 mmol/l
Working reagent
1 ml
3 ml (25,30 a 37C) 300 I
Equilibrate at choosed temperature. Sample Mix and wait 1 mn. 100 l
Measure the extintion decrease per minute for 1-3 minutes. CALCULATION 340 nm LINEARITY If the OD/mn. at 340 nm is greater than 0.15 repeat the test using a sample dilluted 1:10 with saline solution. Multiply result by 10. REFERENCES VALUES 25C Women Men Up to 16 U/l Up to 22 U/l 30C Up to 22 U/l Up to 29 U/l 37C Up to 31 U/l Up to 40 U/l ALAT (U/L) = OD/mn x 1750
PREPARATION AND STABILITY Reconstitute one vial of reagent 2 with the appropriate volume of buffer reagent 1 This working reagent is stable 7 days at 2 - 8C or 24 hours at 20-25C. SAMPLES Serum, heparinized plasma free of hemolysis. PROCEDURE Wavelength : 340 nm 25-30-37C Temperature : Cuvette : 1 cm light path Zero adjustment : air or distilled water.
NOTES Heamolysis will interfere. BIBLIOGRAPHY Bergmeyer, H. Schalbe and Walefeld. Clin. Chem. 24,53-73 (1978). Bergmeyer and Horder Clin. Chem. Acta 105 147 F (1980).
FT An 07
Oct 2006
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PRESENTATION
Ref. 20015 (60 Tests) R1 : 20 x 3 ml R2 : 1 x 7 ml Ref. 20016 (100 Tests) R1 : 10 x 10 ml R2 : 1 x 11 ml Ref. 20017 (120 Tests) R1 : 4 x 30 ml R2 : 2 x 7 ml Ref. 20018 (250 Tests) R1 : 5 x 50 ml R2 : 2 x 14 ml
ALKALIN PHOSPHATASE
ALP (DGKC)
CALCULATION
PRINCIPE Determination of alkaline phosphatase in serum or plasma conforming to the recommandations of deutsche gesellschaff for clinical chemistry. ALP P-nitrophenyl phosphate H2O ---> 4-nitrophenol + phosphate REAGENT R1 DEA Diethanolamine buffer pH 9.8 (37C) MgCI2 R2 substrate Sodium p-nitrophenyl phosphate (PNPP) 1mol/I
Calculate the average value of variation of absorbance per minute (OD/mn) and obtain the enzymatic activity value of the sample, using the following formula : 405 nm ALP (U/L) = OD/mn x 2750 410 nm ALP (U/L) = OD/mn x 2910 LINEARITY If the OD /mn is greater than 0.250 repeat the test using a sample diluted with saline solution and multiply result by the diluted factor. REFERENCE VALUES
0.5 mmol/l
10 mmol/I 25C 30C 37C
SAMPLE Serum, or heparinized plasma, do not use hemolysed samples. PREPARATION OF WORKING SOLUTIONS Ref 20015 R1 3 ml R2 0,3 ml Ref 20016 R1 10 ml R2 1 ml Ref 20017 R1 30 ml R2 3 ml Ref 20018 R1 50 ml R2 5 ml STORAGE AND STABILITY The expiry date of each reagent stored at 2 to 8C is printed on the label. The working solutlon is stable for 15 days at +2 to +8C and for 5 days at 15 to 25C if stored protected from direct light. PROCEDURE Wavelength : Cuvette : Temperature : Measurement :
Children Adults
up to 400 U/l up to 500 U/l up to 650 U/l 40 -190 U/l 50 - 230 U/l 70-300 U/l
Anyway each laboratory should assign their own reference values. REFERENCES 1. Hausamen J.U., Helger R, Rick W.Gross W . Clin. Chim. Acta 15,241 (1967) 2. Bessey O A. Lowry O H Brock M.J J.Biol.Chem 164,321 (1947) 3. Committee of the Scand.Soc for Clin.Chem. Scand.J.Clin.Lab Invest. 33,291 (1974)
405 nm (Hg-400-Hg 420) 1 cm light path 25, 30, 37C against air or distilled water
The working solution must be brought at the choosed temperature for the analysis before use. Pipette directly Working solution Equilibrate (25, 30, 37C) Sample 1ml
20 l
Mix and wait for 1 minute. Read the initial absorbance and start timer immediately. Read again at constant intervals for 3 minutes. 17 FT An 08 Oct 2006
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PRESENTATION
Ref. 20039 200 Tests GOT R1 : 2 x 100ml R3 : 2 x 100ml R4 : 1 x 14ml Ref. 20041 Ref. 20048 50 Tests GOT+ 50 Tests GPT 100 Tests GPT R1 : 1 x 50ml R2 : 2 x 50ml R2 : 1 x 50ml R3 : 2 x 50ml R3 :1 x 100ml R4 : 1 x 9ml R4 : 1 x 9ml Ref. 20049 100 Tests GOT R1 : 2 x 50m1 R3 : 2 x 50ml R4 : 1 x 9ml
AMINO TRANSFERASES
(GOT-GPT) Colorimetric Method
GOT units/ml GPT Units/ml 0 0 22 25 55 50 95 83 150 126
PRINCIPLE
Colorimetric determination of GOT or GPT activity according to the following reactions: GOT L. Aspartate + ketoglutatate ----> Oxaloacetic + L. glutamate GPT
Alanine +Ketoglutatate ----------> pyruvate + glutamate
GPT : Alanine + a-ketoglutate GPT pyruvate + glutamate The pyruvate or oxaloacetate formed is measured in its derivated form, 2.4-dinitrophenylhydrazone.
Plot the standard curve : numberof units/ml in x axis OD (millimetre paper) or percentages of transmission (semi-log paper) in y axis. 2- Measurement Set up the following tubes for each serum:
REAGENTS
Reagent 1 GOT substrate Phosphate buffer pH 7,5 Aspartate -ketoglutarate Phosphate buffer pH 7.5 alanine a-ketoglutarate 2.4 dinitrophenylhydrazine 1 mmol/l Standard pyruvate 85 mmol/l 200 mmol/l 2 mmol/l
GOT Reagent 1 1 ml -
G PT 1 ml
Reagent 2 GPT substrate
95 mmol/l 200 mmol/l 2 mmol/l
Reagent 2
Incubate for 5 minutes at 37C. Serum Mix and incubate at 37C for: Reagent 3 0.2 ml exactly 1 hour 1 ml 0.2 ml exactly 30 mn. 1 ml
Reagent 3 Colour reagent Reagent 4 standard
Mix. Let stand for 20 minutes at room temperature.
STABILITY
The reagents are stable several years at 2-8C. Reagent 3 should be stored in the dark. NaOH 0.4 N subsidiary reagent.
NaOH 0.4 N
10 ml
10 ml
Mix. Wait 5 minutes, Measure under conditions identical to those used for the standard curve. The color intensity is stable during 1 hour
SAMPLES
Serum Heamolysis will interfer.
PROCEDURE
Wavelength: Zero adjustment: 1- Standard curve Pipette into test tubes (ml) : Tube no Distilled water Reagent 1 Reagent 4 Reagent 3 1 0.2 1 1 2 0.2 0.9 0.1 1 3 0.2 0.8 0.2 1 4 0.2 0.7 0.3 1 5 0.2 0.6 0.4 1 505 nm (490 - 520 nm) distilled water
RESULTS
Calculate the number fot GOT and GPT unitis/ml of serum using the standard curve.
LINEARITY
For GOT activities > 150 units/ml and GPT activities > 125 untis/ml re-measure after diluting the sample 1:10 in a 9 g/l sodium chloride solution. Multiply the result by 10
REFERENCE VALUES
GOT : < 40 units/ml GPT : < 45 units/ml
Mix. Let stand for 20 minutes at room temperature. NaOH 0.4 N 10 10 10 10 10
BIBLIOGRAPHY
Britman S. Frankel S. - Am. J. Clin. Path. 1657, 28, 56. Cabaud et al. - Am. J. Clin. Path. 1956, 26, 1101. Karmen A. J. Clin. Invest. 34, 131, 1955.
Mix. Wait 5 minutes, Measure. FT An 09 Oct 2006 18
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PRESENTATION
Ref. 20031, kit for 60 Tests ; 20 x 3 ml : ready to use Ref. 20033, kit for 100 Tests ; 10 x 10 ml : ready to use Ref. 20032, kit for 360 Tests ; 30 x 12 ml : ready to use
- AMYLASE
Direct Kinetic colorimetric method CNPG3
PRINCIPLE
CNPG3 + amylase------> CNP + CNPG2 + glucose + maltotriose As shown above, oc amylase hydrolizes the 2 chloro4nitrophenyl -D-maltotrioside (CNPG3) to release 2chloronitrophenol and form 2-chloro-4-nitrophenyl-Dmaltoside (CNPG2) maltotriose (G3) and glucose (G). The rate of formation of the 2-chloro-4-nitrophenol can be detected spectrophotometrically at 405 nm to give a measurement of -amylase activity in the sample.. Pipette into test tube - Amylase Reagent Equilibrate to 37C. Add : Sample : Serum Urine 25 l 10 l 1000 l
REAGENTS
- Amylase CNPG3 Reagent contains 2-Chloro-4-Nitrophenyl--D-maltotrioside Sodium Chloride Calcium Acetate MES Buffer pH 6.0 Potassium Thiocyanate Sodium Azide
Mix and incubate at 37C for 1 minute. Read the initial absorbance and start timer immediatety. Read again at constant intervals for 3 minutes.
CALCULATIONS
2,5 mmol/l 250 mmol/l 4.5 mmol/l 50 mmol/l 375 mmo/l 52 mmol/l In serum : Amylase (U/l) = O D / mn x 3178 In Urine : Amylase (U/l) = O D / mn x 7829
LINEARITY
The method is linear up to 2000 U/L at 37C.
REFERENCE VALUES
37C Serum, plasma Urine < 100 U/l < 380 U/l
PREPARATION STABILITY-STORAGE
Reagent provided is ready to use. Store the reagent refrigerated at 2-8C. The sealed reagent is stable until the expiration date. The reagent is stable for at least 30 days at room temperature (20-25C). Do not use if the absorbance of the reagent is greater than 0.50 The working solution must be brought at +15 to + 25C before use. WARNING: Do not mouth pipette so as to avoid contamination with salivary amylase
BIBLIOGRAPHY
Kaufman RA. Tietz NW. Clin Chem; 26:846. (1980) Ranson. JHC. Curr Prob Surg; 16:1. (1979) Young DS. pestaner LC. Gibberman V. Clin Chem 21: 1 D. (1975).
SAMPLE
Non-haemolyzed serum is the recommended sample. Separate serum from clot after blood collection. Plasma (for heparin tubes may be used, but other anti-coagulants such as EDTA or citrate should be not used) urine
PROCEDURE
Wavelength Cuvette Temperature Measurement Linearity 405 nm 1 cm light path 37C against air or distilled water up to 2000 U/l
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FT An 10
Oct 2006
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PRESENTATION
Ref. 20042 (60 Tests) R1 : 1 x 65 ml R2 : 20 x 3 ml (Lyoph) Ref. 20043 (100 Tests) R1 : 1 x 110 ml R2 : 10 x 10 ml (Lyoph) Ref. 20050 (450 Tests) R1 : 9 x 50 ml R2 : 9 x 50 ml (Lyoph)
ASAT - GOT
Kinetic test IFCC Without Pyridoxal phosphate
PRINCIPLE Kinetic determination of Aspartate amino transferase activity. Method recommended by the IFCC GOT
2-Oxoglutarate + L. Aspartate----------> Glutamate + Oxaloacetate
Working reagent
1 ml
3 ml
Equilibrate at choosed temperature. 25,30 or 37C Sample Mix and wait 1 mn. Measure the extinction decrease per mn. for 1-3 minute. CALCULATION 340 nm ASAT (U/L) = OD / mn x 1750 LINEARITY If the OD/mn. at 340 nm is greater than 0.15 repeat the test using a sample diluted 1 : 10 with saline solution. Multiply result by 10. REFERENCE VALUES 100l 300 l
MDH
Oxalacetate + NADH + H+----------> Malate + NAD+
The rate of NADH consumption is determined photometrically and is directly proportional to the GOT activity in the sample. GOT : Glutamic oxaloactate transaminase MDH : Malate deshydrogenase. REAGENTS
25C Reagent 1 Tris buffer pH 7.8 Buffer reagent L-Aspartate Reagent 2 Substrate NADH LDH MDH Oxoglutarate 80 mmol/I 200 mmol/l 0.18 mmol/l 800 U/l 600 U/l 12 mmol/l Women Men Up to 16 U/l Up to 19 U/I
30C Up to 22 U/I Up to 26 U/I
37C Up to 31 U/l Up to 38 U/I
PREPARATION AND STABILITY Reconstitute one vial of Reagent 2 with the appropriate volume of buffer/Reagent 1 This working reagent is stable : 7 days at + 2- 8C or 24 hours at 20-25C. SAMPLES Serum, heparinized plasma free of heamolysis. PROCEDURE Wavelength : .................................................................. 340 nm Temperature : ....................................................... 25-30-37C Cuvette : ..............................................................1 cm light path Zero adjustment : air or distilled water.
NOTES Hemolysis will interfere. BIBLIOGRAPHY Bergmeyer, M. et al. Clin. Chim. Acta 70, F 19 (1976) Bergmeyer and Wahlefeld Clin. Chem. 24, 58-(1978).
FT An 11
Oct 2006
20
Biomaghreb
PRESENTATION
Rf. 200063, (60 Tests) R1: 2 x 24 ml R2 : 2 x 6 ml Rf 200071, (150 Tests) R1 : 5 x 24 ml R2 : 5 x 6 ml Rf 200072, (180 Tests) R1 : 6 x 24 ml R2 : 6 x 6 ml
CREATINE KINASE LS
NAC activated, IFCC method
Testing procedure
1- Manual procedure : Serum start
Wavelength : Temperature : Cuvette : Zero adjustment : Working reagent Sample 340nm.Hg 334 or Hg 365 nm) +25 / +30 / +37C 1 cm light path air or distilled water Macro 2500l 100l Semi-Micro 1000l 40l Micro 500l 20l
PRINCIPE
Kinetic Dtermination of cratine kinase N. acetyl cystein reactived CK Creatine phosphate + ADP creatine + ATP D- Glucose + ATP HK D-glucose- 6 Phosphate + ADP
G-6-PDH
D-Glucose-6-phosphate- NADP
D-gluconate - 6 -P
+ NADPH+H The catalyte activity of Ck is determined by the measurement of NADPH formation.
Mix and incubate for 2 minutes. Measure the obsorbance increase per minute for 1 - 3 minutes.
Calculation :
340nm A/min.x Hg 334nm A/min.x Hg 365nm A/min.x Macro 4130 4207 7429 Semi-Micro 4130 4207 7429 Micro 4130 4207 7429
REAGENTS
Reagent 1 Tampon imidazole Acetate pH : 6,7 Glucose Reagent 2 : Substrate Cratin phosphate NAC (N. Acetyl cysteine ADP AMP Diadenosine pentaphosphate NADP Hexokinase (HK) G-6- PDH 100 mmol/l 20 mmol/l 30 mmol/l 20 mmol/l 5 mmol/l 2 mmol/l 10 mol/l 2 mmol/l 2500 Ul/l 1500 Ul/l
2- Manual procedure : Substrate start

Wavelength : Temperature cuvette Zero adjustment : R1 Sample R2 340nm.Hg 334 ou Hg 365 nm) +25 / +30 / +37C 1 cm light path air or distilled water Macro 2000l 100l 400l Semi-Micro 800l 40l 160l Micro 400l 20l 80l
Mix and incubate for 2 minutes. Measure the absorbance increase per minute for 1 - 3 minutes.
Calcul
340nm A/min.x Hg 334nm A/min.x Hg 365nm A/min.x Macro 3965 4039 7132 Semi-Micro 3965 4039 7132 Micro 3965 4039 7132
SPECIMEN
Collect serum using standard sampling tubes. Heparinized-or EDTA-plasma Stability : 7 days at +4C to + 8 C 2 days at +20C to + 25 C Centrifuge samples containing precipitate before performing the assay
LINEARITY
Linearity up to 900 U/L at 37C if the OD/min is greater than 0,200 repeat the test using a sample diluted 1 : 10 with saline solution. Multiply result by 10.

1- Serum start : Mix 4 volumes of R1 with 1 volume of R2 This solution is stable up to 10 days at + 2C to + 8C 1 days at + 20C to + 25C Unopened kid components : Up to the expiry date at + 2C to + 8C.
REFERENCE VALUES
Men Women 25C 10-80 U/l 10-70 U/l 30C 15-130 U/l 15-110 U/l 37C 25-195 U/l 25-170 U/l
Anyway each laboratory should assign their own references values.
BIBLIOGRAPHIE
1- Bablok W. et al. A genral Regression Procedure for Method transforma-
2 - Substrate start : R1 : Ready for use R2 : Ready for use Onboard stability : R1 21 days R2 21 days 21
tion JClin Chem Clin Biochem 1988 ; 26 : 78-790 2- Black H.R. Quallich H gareleck CB. Racial diffrences in serum Creatine kinase levels Am J Med 7986 ; 81 : 479-487 3- GLick M.R, Ryder KW, Jackson SA. Graphital Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986 ; 32 ; 470-474. Passing H. Bablok W. A New Biometrical Procedure for Testing the Equality of Measurements from Two Different Analytical Methods. J Clin Chem Clin Biochem 1983 ; 21 : 707-720. 4- Guder W. G., Narayanan S., Wisser H., Zawta B. List of Analytes Preanalytical Variables, Brochure in Samples : From The Patient to the Laboratory. Darnstadt GIT Veriag 1996
FT An 12
Oct 2008
Biomaghreb
PRESENTATION
Ref. 20021 : 60 Tests R1 : 1 x 65 ml R2 : 20 x 3 ml (Lyoph) Ref. 20022 : 100 Tests R1 : 1 x 110 ml R2 : 10 x 10 ml (Lyoph)
GAMMA GT
Kinetic Colorimetric method (I FCC)
PRINCIPLE
Kinetic determination of y-GIutamyl-transferase activity according to the following reaction : -GT Glupa Carboxy+glycylglycine>T L-glutamylplycyl-glycine + 5 amino-2-nitrobenzoate
CALCULATION
At. 405 nm At. 410 nm OD/mn x 1190 = U/l ODImn x 1339= U/L
LINEARITY REAGENTS
Reagent 1 Glycylglycine buffer pH 7,9 150 mmol/l Buffer solution 30C Reagent 2 Glupacarboxy 6 mmol/l Substrate This method is linear at 500 u/l at 37C If the AOD/min is greater than 0.200 repeat the test using a sample diluted 1 : 10 with saline solution multiply result by 10.
REFERENCE VALUES
25C Men Women 6-28 U/l 4-18U/I 30C 8-38 U/l 5-25 U/l 37C 11-50 U/l 7-32 U/l

Reconstitute one vial of Reagent 2 with the appropriate volume of buffer (Reagent 1 ) This working reagent is stable : 7 days a 20-25C 1 month at +4C
SAMPLES
Serum free of hemolysis.
NOTES
Citrated, oxalated and EDTA-plasma can not be used in this test. 405 nm (410) 25-30-37C 1 cm light path
PROCEDURE
Wavelength : Temperature : Cuvette : Zero adjustment : air or distilied water. Working reagent 1 ml 3 ml
BIBLIOGRAPHY
Lum, G. and Gabino, SR. Clin. Chem. 18,358 (1972) Szasz, G. Clin. Chem. 15,124 (1969).
Equilibrate at choosed temperature (25,30 or 37C) Sample Mix and wait 1 mn. Measure the extinction increase per minute for 1-3 minutes. 100 I 300l
FT An 13
Oct 2006
22
Biomaghreb
PRESENTATION
Ref. 20011 (60Tests) R1 : 1 x 65 ml R2 : 20 x 3 ml (Lyoph) Ref. 20012 (100Tests) R1 : 1 x 110 ml R2 : 10 x 10 ml Iyoph Ref. 200123(450Tests) R1 : 9 x 50 ml R2 : 9 x 50 ml Iyoph Ref. 200125(150Tests) R1 : 1 x 160 ml R2 : 15 x 10 ml
LDH (SFBC)
Kinetic method
CALCULATION
340 nm OD /mn.x 8095 = U/l
PRINCIPLE
Kinetic determination of the Lactate dehydrogenase activity optimized test according to the recommendation of SFBC (societe franaise de biologie clinique) LDH Pyruvate + NADH + H+ Lactate + NAD+
LINEARITY
If the OD/mn at 340 is greater than 0.160, repeat the test using a sample diluted 1 : 10 with saline solution Multiply result by 10.
The activity of LDH is shown by the variation of OD at 340 nm which is proportional to the quantity of NADH oxidised.
REFERENCES VALUES
30C 140 - 280 U/l 37C 230- 430 U/l
Adults
REAGENTS
Reagent 1 This buffer pH 7.2 at 30C 80 mmol/l Buffer reagent Pyruvate 1.6 mmol/l Nacl 200 mmol/I Reagent 2 Coenzyme NADH 0.2 mmol/l Children to 0 at 6 months 415 - 690 U/l 565 - 940 U/l
NOTES
Heamolysis will interfere.

Reconstitute one vial of Reagent 2 with the appropriate volume of buffer / Reagent 1. This working reagent is stable : 15 days at 2-8C or 24 hours at 20-25C.
BIBLIOGRAPHY
Bergmeyer, H.U., J. Clin. Chem. Glin Biochem. 13,507 (1975) Howell B.F. and coll. Clin. Chem. 25, 269 (197g). Commission Enzymologie - SFBC - Inform Sci - Biol - 1981,5.
SAMPLES
Serum or heparized plasma without hemolysis.
PROCEDURE
Wavelength : Temperature : Cuvette : 340 nm 30-37C 1 cm light path
Zero adjustement : air or distilled water. Working Reagent Equilibrate at choosed temperature Sample Mix and wait 1 mn Measure the extinction decrease per mn. for 1-3 20 l 60 l 1 ml 3 ml
23
FT An 14
Oct 2006
Biomaghreb
BIOCHEMICAL SUBSTRATES
Page
ALBUMIN
Ref. 20094 4 x 250 ml
26
CALIBRATOR FOR TOTAL AND DIRECT BILIRUBIN

Ref. 201010 10 x 3 ml
27 28
CHOLESTEROL
CHOLESTEROL HDL
Ref. 20113 Ref. 200113 Ref. 200112 5 x 5 ml 10 x10 ml 1 x 5 ml
29
CREATININE
30
DIRECT BILIRUBIN
Rf. 20102 2 x 160 ml
31 32 Ref. Ref. Ref. Ref. Ref. 20121 20126 20124 20122 20127 2 x 500 5 x 200 6 x 500 3 x 1000 4 x 100 ml ml ml ml ml 33 2 x 160 ml 2 x 600 ml 2 x 250 ml 34 Ref. 20103 2 x 160 ml 35 Ref. 20161 Ref. 20162 450 ml 1000 ml 36 Ref. 20131 Ref. 20132 Ref. 20138 2 x120 ml 4 x 30 ml 5 x120 ml 37 2 x100 ml 1 x 500 ml 2 x 500 ml 38 4 x50 ml 9 x50 ml 39 Ref. 20091 Ref. 20092 Ref. 20095 3 x125 ml 4 x30 ml 2 x120 ml
GLUCOSE
TOTAL AND DIRECT BILIRUBIN

Ref. 20101 Ref. 20104 Ref. 20105
TOTAL BILIRUBIN TOTAL PROTEINS TRIGLYCERIDES
UREA (COLORIMETRIC METHOD)

Ref. 20141 Ref. 20146 Ref. 20148
UREA (KINETIC METHOD)

Ref. 20143 Ref. 20147
URIC ACID
25
Biomaghreb
PRESENTATION
Ref. 20094 (500 tests) R1 : 4 x 250 ml R2 : 2 x 3ml
ALBUMIN
BCG Method
PRINCIPLE
Albumin, in a buffered solution, reacts with the green of bromocresol (BCG) to form a red-colour complex
CALCULATION
OD Sample OD Standard g/l : n = 50 mol/l : n = 724,5 xn
REAGENTS
Reagent 1 : Bromocresol green Succinate Buffer Brij Bovine Albumine 0,14 g/l 75 mml/l 7ml/l 50 g/l 724ml/l
Reagent 2 : Standard
LINEARITY
This method is linear up to 1000 mol/l (69 g/l)
REFERENCE VALUES STORAGE AND STABILITY

The expiry date of reagent stored at 2-8C is printed on the label Serum / Plasma 550-780 mol/l 38 - 54 g/l
SAMPLES
Serum, heparinized plasma
BIBLIOGRAPHIE
Doumas B et al.clin. chim Acta 31,87 (1971). Drupt, F. Pharm. Biol 9.777 (1974) 628 mm 20 - 25C 1 cm light path
PROCEDURE
Wavelength : Temperature : Cuvette :
Read against blank reagent, standard and sample. Prepare a standard and a blank for each series of determinations the BCG reagent must be brought at 15 - 25C before use. Pipette into tests tubes Blank 2 ml Standard 10 l 2 ml Sampie 10l 2 ml
Sample Standard Reagent 1
Mix and read after 5 minute the OD of sample and standard against blank the colour is stable for 30 minutes
FT An 15
Oct 2006
26
Biomaghreb
CALIBRATOR FOR TOTAL AND DIRECT BILIRUBIN
PRESENTATION
Ref. 201010 10 x 3 ml (Iypophilized) The calibrator containing conjugated and unconjugated bilirubin in a matrix of bovin albumin. The exact concentrations are given on each vial.
CALIBRATEUR BILIRUBINE TOTALE ET DIRECTE

PRESENTATION
Ref. 201010 10 x 3 ml (Iyophilise) Le calibrateur est constitue de bilirubine conjuguee et de bilirubine non conjuguee dans une matrice dalbumine bovine. Les concentrations exactes sont indiquee sur chaque flacon.
USE
Calibrator of the determination of total and direct bilirubin of the following technique: - Total Bilirubin (Ref : 20103) - Direct Bilirubin (Ref : 20102) - Total and direct Bilirubin (Ref : 20101)
UTILISATION
Calibrage du dosage de la bilirubine totale et directe pour les techniques - Bilirubine totale (Ref : 20 103) - Bilirubine directe (Ref : 20 102) - Bilirubine totale et directe (Ref : 20 101 )
Reconstitution
Open a vial carefully, avoinding any loss of the Iypophilized material. Reconstitute with exactly 3 ml of distilled water. Let stand for 15 minutes and then dissolve the contents completely by gently inverting the vial (do not shake).
Reconstitution
Ouvrir avec precaution un flacon sans perte de Iyophilisat le reprendre par exactement 3 ml deau distillee. Attendre 15 minutes completer la dissolution par retournement successifs de flacon (ne pas agiter).
PRECAUTIONS
No known analysis method can totelly guarantee the absence of transmissible pathogenic agents. It is therefore recommended that these products be treated as potentially infections, and handled observing the usual safety precaution.
PRECAUTION DUTILISATION
Aucune des methodes danalyse actuellement connues ne peut garantir de fa,con absolue que les produits ne contiennent aucun agent pathogane transmissible.ll est recommande de les manipuler avec les precautions dusage relatives au produit potentiellement infectieux.
STABILITY
Lyophilized form Stable at 2-8c until expiration date given on the packaging.
STABILITE
Forme Iyophilisee Ce produit est stable a 2-8c jusqua la date de peremptions indiquee sur chaque conditionnement.
After reconstitution, in the dark: - 2 days at 20-25c - 4 days at 2-8c - 6 weeks at -20c
Apres reconstitution a lobscurite - 2 jours a 20-25c - 4 jours a 2-8c - 6 semaines a - 20c
27
FT An 16
Oct 2006
Biomaghreb
PRESENTATION
Ref. 20111(360Tests) R1 : 3 x 120 ml R2 : 3 Vials (Iyophilized) R3 : 1 x 5ml Ref. 20112 (120Tests) R1 : 4 x 30ml R2 : 4 Vials (Iyophilized) R3 : 1 x 4ml Ref. 20115 (600Tests) R1 : 6 x 100ml R2 : 6 Vials (Iyophilized) R3 : 2 x 5ml
CHOLESTEROL
Enzymatic Colorimetric test (CHOD-PAP)
PRINCIPLE
The cholesterol is determined after enzymatic hydrolysis and oxidation. The indicator quinoneimine is formed from hydrogen peroxide and 4-aminoantipyrine in the presence of phenol and peroxidase.
Cholesterol esterase Esters of cholesterol + H2O-------> Cholesterol + Fatty acids Cholesterol oxidase Cholesterol + O2 --------------------> Cholest-4-en-one + H202 Peroxidase H2O + Phenol + 4Aminophenazone ----------> Quinonimine
Pipette into tests tubes Blank 1 ml Standard 10 l 1ml Sample 10 l 1 ml
Standard Sample Working reagent
Mix. incubate 5 mn. at 37C before reading. The colour is stable for 30 mn.
CALCULATION
OD sample Cholesterol = OD standard mg/dl: g/l: mmol/l: n = 200 n=2 n = 5.17 xn
The quantity of this red dye quinonimine formed is proportional to the cholesterol concentration.
REAGENTS
Reagent 1 Buffer solution Pipes pH 6.9 Phenol 90 mmol/l 26 mmol/l
LINEARITY
This method is linear up to 600 mg/dl (15.4 mmol/l). If the cholesterol concentration is greater than 600 mg/dl (6 g/l) in the serurn or plasma, dilute the sample 1 : 2 with saline solution and repeat the determination and multiply the result by 2.
Reagent 2 vial of enzymes
Cholesterol oxidase 300 U/l Peroxidase 1250 U/l Cholesterol esterase 300 U/l 4-Aminophenazone 0.4 mmol/l 200 mg/dl 2 g/l 5.17 mmol/l
Reagent 3 standard
REFERENCE VALUES
Serum or plasma : 3.6 - 7 mmol/l 1.4 - 2.7 g/l 140 - 270 mg/dl 260 mg/dl (6.7 mmol/l 2,60 g/l)

Reconstitute one vial of reagent 2 with the approriate volume of buffer/reagent 1 This working reagent is stable 4 months at 2 - 8C or 1 month at 20 - 25C.
Increased risk above :
BIBLIOGRAPHY
Fasce, CF, Clin. Chem. 18, 901 (1982) Richmond, Clin. Chem. 19, 1350 (1973) Trinder P. Ann. Clin. Biochem 16.24 (1969)
SAMPLES
Serum, heparinized piasma
PROCEDURE
Wavelength: Temperature: Cuvette: 505 nm (500-550) 37C 1 cm lightpath
Read against blank reagent, standard and sample.
FT An 17
Oct 2006
28
Biomaghreb
PRESENTATION
Ref. 20113 (500 Tests) 5 x 5ml Ref. 200112 (100 Tests) 1 x 5ml Ref. 200113 (2000 Tests) 10 x 10ml
CHOLESTEROL- HDL
Pipette into the tubes : Blank Distilled water Standard (2 g/l) Supernatant Reagent solution cholesterol assay 10l 1 ml Standard Sample 10l 1 ml 10l 1 ml
Auxiliary kit Ref : 20111 Ref : 20112 Ref : 20115 PRINCIPLE Low density lipoproteins (LDL and VLDL) and chylomicron fractions are precipitated quantitatively by the addition of phosphotungstic acid in the presence of magnesium ions. After centrifugation, the cholesterol concentration in the HDL thigh density lipoprotein) fraction, which remains in the supernatant, is determined using. Cholesterol kit (Ref : 20111 -20112).
Mix, incubate for 5 minutes at 37C. Read against the reagent blank within 30 minutes.
REAGENT
Phosphotungstic Acid Magnesium chloride pH 6,2 13,9 mmol/l 490 mmol/l
CALCULATION
OD measurement
[HDL-Cholesterol] =
OD standard n = 5.17 mmol/l n = 2 g/l
xn
STABILITY
Stable up the expiry date specified when stored at 2 - 8C. Discared any troubled or coloured reagent.
Multiplie the result per 1.1 (1/11 is the dilution with the precipitant reageant) we obtaint the concentration of cholesterol bound to HDL N.B : once the bottle of reagent HDL is started, there is a possibility of formation of the brillant crystals at the bottom of the bottle which havent any influences on the quality of the product
SAMPLE
Serum plasma collected on EDTA
PROCEDURE
Dilute in NaCI 9g/l solution, serum with triglycerides more than 3,5 mmol/l. The supenatant may be stored up to five days at 2 - 8C
INTERPRETATION
European Athero sclerosis Society has established relationship between risk level of cornar diseases and cholesterolemia. Cholesterolemia < 2 g/l < 5,2 mmol/l 2,0 to 2,5 g/l 5,2 to 6,5 mmol/l > 2,5 g/l > 6,5 mmol/l Risk level Low risk
Precipitation
Serum 500 l 50 l Precipitant Reagent mix and allow for 10 mn then centrifuge at 5000 rpm for 15 mn.
Moderate risk if
Cholesterol CHOD-PAP Assay

Reagent Solution for Cholesterol Assay : See pack insert of Chotesterol, CHOD-PAP (Ref 20 111 or 20 112) Wavelenght : Cuvette : Temperature : Measurement : 500 nm, (492-550 nm) 1 cm light path 37C against reagent blank
HDL Cholesterol < 0,35 g/l < 0,9 mmol/l)
High risk if
REFERENCES
ARCOL.ISB - 15, 121 - 124 (1989) Burstein M et al. Lipid Res 11, 583 (1970). Study group .European Atherosclerosis European heart journal 1988. Society,
29
FT An 18
Janvier 2008
Biomaghreb
PRESENTATION
Ref. 20151 (320 Tests) R1 : 2 x 80ml R2 : 2 x 80 ml R3 : 1 x 15 ml Ref. 20152 (3000 Tests) Ref. 20153 (1000 Tests) R1 : 3 x 500ml R1 : 1 x 500ml R2 : 3 x 500 ml R2 : 1 x 500 ml R3 : 3 x 50 ml R3 : 2 x 25 ml
CREATININE
Kinetic test without deproteinization
CALCULATION
PRINCIPLE Creatinine in a basic picrate solution forms a colored complex. The extinction at predetermined times during conversion is proportional to the concentration of creatinine in the sample. REAGENTS Reagent 1 Reagent 2 Reagent 3 Sodium hydroxide Picric acid solution Standard Creatinine 1.6 mol/l 17.5 mmol/l 2 mg/dl 20 mg/l 176.8 mol/l
Having calculated the OD=OD2-OD1. The samples creatinine concentration will be obtained using the following formula. OD Sample [Creatinine] = xn OD Standard mg/dl : n = 2 mg/l : n = 20 mol/l : n = 176.8 LINEARITY This method is linear up to 15 mg/dl (1326 mol/l - 150 mg/l). If the creatinine concentration is greater than 15 mg/dl in the serum, dilute the sample 1/2 with saline solution NaCl 9g/l and repeat the determination and multiply the result by 2. REFERENCES VALUES Serum 0.7 - 1.4 mg/dl 7 - 14 mg/l 61.8 - 132.6 mol/l Urine 15 - 25 mg/kg/24 h
PREPARATION AND STABILITY The reagents are ready for use and stable at room temperature, up to the date of expiration as specified. WORKING REAGENT Mix proportionaly 1 : 1 ; the reagent R1 and R2 Stability : 1 month at 20-25C SAMPLES Serum, heparinized plasma Urine diluted 1/20 PROCEDURE Wavelenght : Temperature : Cuvette : Read against air or distilled water. Pipette into the tubes Standard Standard Sample Working Reagent 100 l 1 ml Sample 100 l 1 ml
492 nm (490-510) 25, 30 ou 37C 1 cm lightpath
BIBLIOGRAPHY Henry, J.B. Clinical Diagnosis and management 17 Th edition, Sauders Publisher (1984). Larsen, K. Clin. Chim Acta 66, 209 (1972).
Mix and read OD1 after 30 sec. Read again OD2 exactely after 1 mn.
FT An 19
Oct 2006
30
Biomaghreb
PRESENTATION
Ref. 20102 (160 Tests) R2 : 4x80ml R3 : 1 x 21 ml R4 : 1 x 3 ml (Iyophilized)
DIRECT BILIRUBIN
PRINCIPLE
Sulfanilic acid reacts with sodium nitrite to form diazotized sulfanilic acid. In the absence of Dimethylsulfoxide, only direct bilirubin reacts with diazotized sulfanilic acid to form azobilirubin.
Standard Blank Standard R4 Sample Reagent R2 Working solution 50 l 1 ml 1 ml Reac 50 l
Sample Blank Reac
REAGENTS
Reagent2 Reagent 3 Reagent 4 Sulfanilic acid Hydrochloric acid Sodium nitrite Standard 30 mmol/l 150 mmol/l 20 mmol/l
50 l 1 ml
50 l 1 ml
Mix well and incubate exactly 5 minutes at 37C. Read the absorbance (A) of standard and Sample against their blank.
SAMPLES
Serum or heparinized plasma or collected or EDTA Citrate or Fluoride non hemolysed - stored on protection of ligth.
CALCULATION (DB)
Abs (A) sample [Direct Bilirubin] = Abs ((A))standard [Direct bilirubin] : Abs (A) sample x Factor Note : x 0.585 mol/l x 1.710 mg/l x [std conc]
STANDARD PREPARATION
Reconstitute contents of R4 vial with exactly 3 ml of distilled water. Let stend for 15 minutes and than dissolve the contents completely by gently inverting the vial (do not shake). The exact concentrations are given on each vial. Stability after reconstitution, in the dark : - 2 days at 20-25C - 4 days at 2-8C - 6 weeks at - 20C its necessary to obtain calibration factor on each own laboratory conditions. Std Direct Bilirubin concentration F= Abs standard - Abs std blank
LINEARITY
BD: Linear up to 100 mg/l (10 mg/dl -170 mol/l).
REFERENCE VALUES
Serum < 0.2 mg/dl < 2 mg/l < 3.4 mol/l
PROCEDURE
Working solution Mix 20 R2 vol with 1 R3 vol Wavelength : 555 nm (546 Hg). Temperature : 37C 1 cm light path. Cuvette : Measurement : Against Standard blank for Standard Against Sample blank for sample Stability in the dark 6 hours at 20-25C 2 days at 2-8C
BIBLIOGRAPHY
Hijmans Van den Bergh A.A., Muiler P., Biochem. 77,90 (1916) Walters M.l., Gerarde R.W., Microchem. 15, 23 (1970)
31
FT An 20
Oct 2006
Biomaghreb
PRESENTATION
Ref. 20121: (1000tests) R1 : 2 x 500ml . R2 : 2 vials (Iyoph) R3 : 2 x 6 ml Ref. 20122 : (3000tests) R1 : 3 x 1000ml R2 : 3 vials (Iyoph) R3 : 3 x 11 ml Ref. 20124 : (3000 tests) R1 : 6 x 500ml R2 : 6 vials (Iyoph) R3 : 3 x 11 ml Ref. 20126 : (1000 tests) R1 : 5 x 200ml R2 : 5 vials (Iyoph) R3 : 2 x 6ml Ref. 20127: (400 tests) R1 : 4 x 100ml R2 : 4 vials (Iyoph) R3 : 1 x 5ml
GLUCOSE
Enzymatic Colorimetric (GOD-PAP)
CALCULATION O.D.Sample Glucose conc. = xn O.D.Standard mg/dl : n = 100 g/l : n = 1 mmol/l : n = 5.56 LINEARITY This method is linear up to 500 mg/dl (27.8 mmol/l - 5 g/l). If the glucose concentration is greater than 500 mg/dl repeat the determination using a sample diluted 1/2 with saline solution and multiply the result by 2. REFERENCES VALUES Serum, plasma 70-105 mg/dl 3.89 - 5.84 mmol/l 0.70 - 1.05 g/l 50-70 mg/dl 2.78 - 3.89 mmol/l 0.50 - 0.70 g/l
PRINCIPLE Glucose is oxidized by glucose-oxidase to gluconate and hydrogene peroxide according to the following equation. GOD Glucose + O2 + H2O -------> H2O2 + Gluconate
Peroxidase 2H2O2 + Phenol + 4 Amino-antipyrine ---> 4H2O + Quinonimine
REAGENTS Reagent 1 Buffer solution Reagent 2 Tris buffer pH 7 Phenol Glucose oxidase Peroxidase 4-amino-antipyrine Standard glucose 100 mmol/l 0.3 mmol/l 10 000 U/l 1 000 U/l 2.6 mmol/I 100 mg/dl 5.56 mmol/l 1g/l
Reagent 3 Standard
Spinal fluid
PREPARATION AND STABILITY Dissolve each vial (R2) with buffer (R1), (in brown vial): stability 8 weeks at 20-25C 8 months at 2-8C. SAMPLES Serum (not hemolyzed). Plasma collected on heparin fluoride or heparin iodoacetate (not hemolyzed) Spinal fluid. PROCEDURE Wavelenght : Temperature : Cuvette : Zero adjustment : blank reagent Blank Standard Sample Working reagent 1 ml
NOTES Do not interfere : Hemoglobine (4 g/l), Bilirubine (200 mg/l). Creatinine (100 mg/l), Galactose (1 g/l) and EDTA (2 g/l) BIBLIOGRAPHY Dingeon, B. Ann. Biol. Clin. 33,3 (1g75) Lott, J.A. Clin. Chem. 21, 1754 (1975) Trinder, P. Ann. Clin. Biochem. 6,24 (1969)
505 nm (490-550) (20, 25, 37C) 1 cm lightpath
Standard 10 l 1 ml
Sample 10 l 1 ml
Mix. incubate 10 mn. at 37C or 30 mn at room temperature (20C - 25C). The colour is stable 30 mn.
FT An 21
Juin 2008
32
Biomaghreb
PRESENTATION
Ref. 20101 (160 Tests) R1 : 2 x 80ml R2 : 2 x 80 ml R3 : 1 x 21 ml R4 : 1 x 3 ml (Iyophilized) Ref. 20104 (600 Tests) Ref. 20105 (250 Tests) R1 : 6 x 100ml R2 : 6 x 100 ml R3 : 2 x 40ml R1 : 2 x 125ml R2 : 2 x 125 ml R3 : 1 x 20ml
TOTAL AND DIRECT BILIRUBIN

Standard Blank Reac 50 l 50 l Sample Blank Reac
R4 : 4 x 3 ml (Iyophilized) R4 : 2 x 3 ml (Iyophilized)
PRINCIPLE
Sulfanilic acid reacts with sodium nitrite to form diazotized sulfanilic acid. In the presence of Dimethyl sulfoxide, total bilirubin reacts with diazotized sulfanilic acid to form azobilirubin. In the absence of Dimethylsulfoxide, only direct biiirurbin reacts with diazotized sulfalinic acid to form azobilirubin.
Standard R4 Sample 50 l 50 l Reagent R1 1 ml 1 ml Working solutiom 1 ml 1 ml Mix well and incubate exactly 5 minutes at 37C. Read the absorbance (A) of standard and sample against their blank.
REAGENTS
Reagent 1 Sulfanilic acid Hydrochloric acid Dimethylsulfoxide Sulfanilic acid Hydrochloric acid Sodium nitrite Standard 30 mmol/l 150 mmol/l 7 mol/l 30 mmol/l 150 mmol/l 20 mmol/l See. Standard preparation
N.B : To dilute the samples of new-born or very icteric to the 1/5, in a pysiologic solution (9g / l of Nacl).
Direct Bilirubin Mix 20 volumes R2 with 1 vol R3 Stability in the dark 6 hours at 20-25C 2 daysat+2-8C Standard Blank Reac 50 l 50 I Test Blank Reac
Reagent2
Reagent 3 Reagent 4
SAMPLES
Serum or heparinized plasma or collected or EDTA Citrate or Fluoride non hemolysed stored in the dark.
PROGEDURE
1- Standard preparation Reconstitute contents of R4 vial with exactly 3 ml of distilled water. Let stend for 15 minutes and than dissolve the contents completely by gently inverting the viai (do not shake). The exact concentrations are given on each vial. Stability after reconstitution, in the dark: - 2 days at 20-25C - 4 days at 2-8C - 6 weeks at - 20C its necessary to obtain calibration factor on each own laboratory conditions. F= std Total or sid Direct Bilirubin concentration Abs standard - Abs std blank
Standard R4 Sample 50 l 50l Reagent 2 (D) 1 ml 1 ml Working solution (D) 1ml 1 ml Mix well and incubate exactly 5 minutes at 37C. Read the absorbance (A) of standard and Test agains their blank.
CALCULATION (TB and DB)

[Total or Direct Bilirubin] = Abs (A)satandard x [std T or D conc]
Abs (A) sample
or [Total or Direct bilirubin] : Abs (A) sample x Factor Note: x 0.585 mol/l x 1.710 mg/l
LINEARITY
BT : Linear up to 200 mg/l (20 mg/dl - 340 mol/l) BD : Linear up to 100 mg/l (10 mg/dl -170 mol/l).
2- Reading 555 nm (546 Hg). Wavelength : Temperature : 37C Cuvette : 1 cm light path. Measurement : Against standard blank for standard Against sample blank for sample 3- Working solution - Total Bilirubin Mix 20 R1 Vol with 1 R3 Vol Stability in the dark 6 hours at 20-25C 2 days at + 2-8C 33
EXPECTED VALUES
Total Bilirubin : 0.2 - 1.0 mg/dl (2-10 mg/l ; 3.4 - 17 mol/l) Direct Bilirubin : 0.0 - 0.2 mg/dl ; 0.0 - 2 mg/I ; (0.0 - 3.4 mol/l).
BIBLIOGRAPHY
Hijmans Van den Bergh A.A., Muller P., Biochem. J. 77,90 (1916) Walters M.l., Gerarde R.W., Microchem. J.15, 231 (1970) FT An 22 Juin 2008
Biomaghreb
PRESENTATION
Ref. 20103 (160 Tests) R1 : 4 x 80ml R3 : 1 x 21 ml R4 : 1 x 3 ml (lyophilized)
TOTAL BILIRUBIN
PRINCIPLE
Sulfanilic acid reacts with sodium nitrite to form diazotized sulfanilic acid. In the presence of Dimethyl sulfoxide, total bilirubin reacts with diazotized sulfanilic acid to form azobilirubin.
REAGENTS
Reagent 1 Sulfanilic and Hydrochloric acid Dimethylsulfoxide Sodium nitrite Sandard 30 mmol/l 150 mmol/l 7 mol/l 20 mmol/l
Standard R4 Sample 50 l 50 l Reagent R1 1 ml 1 ml Working solutiom 1 ml 1 ml Mix well and incubate exactly 5 minutes at 37C. Read the absorbance (A) of standard and sample against their blank.
Standard Blank Reac 50 l 50 l
Sample Blank Reac
Reagent3 Reagent 4
N.B : To dilute the samples of new-born or very icteric to the 1/5, in a pysiologic solution (9g / l of Nacl). CALCULATION (TB)
[Total Bilirubin] = or [Total bilirubin] : Abs (A) sample x Factor Note: x 0.585 mol/l x 1.710 mg/l Abs (A) sample Abs (A)satandard x [std conc]
SAMPLES
Serum or heparinized plasma or collected or EDTA Citrate ou Fluoride non hemolysed - stored on protection of ligth.
STANDARD PREPARATION
Reconstitute contents of R4 vial with exactly 3 ml of distilled water. Let stend for 15 minutes and than dissolve the contents completely by gently inverting the vial (do not shake). The exact concentrations are given on each vial. Stability after reconstitution, in the dark: - 2 days at 20-25C - 4 days at 2-8C - 6 weeks at - 20C its necessary to obtain calibration factor on each own laboratory conditions. F= std Total Bilirubin concentration Abs standard - Abs std blank
LINEARITY
BT : Linear up to 200 mg/l (20 mg/dl-340 mol).
REFERENCE VALUES
Serum Total Bilirubin < 1 mg/dl < 10 mg/l < 17 mol/l
PROCEDURE
Working solution Mix 20 R1 vol with 1 R3 vol Wavelength : 555 nm (546 Hg). Temperature : 37C Cuvette: 1 cm light path. Measurement : - Against standard blank for standard - Against sample blank for sample. Stability in the dark 6 hours at 20-25C 2 days at + 2-8C
BIBLIOGRAPHY
Hijmans Van den Bergh A.A., Muller P., Biochem. 77,90 (1916) Walters M.l., Gerarde R.W., Microchem. 15, 23 (1970)
FT An 23
Juin 2008
34
Biomaghreb
PRESENTATION
Ref.20161 (90 to 450 Tests) R1 : 3 x 147ml R2 : 1 x 11ml R3 : 1 x 4,5 ml Ref.20162 (200 to 1000) Tests R1 : 4 X 245 ml R2 : 1 x 25 ml R3 : 2 x 4 ml
TOTAL PROTEINS
Colorimetric test (Biuret)
PRINCIPE
Serumal proteins formed in alkaline medium a coloured complex whose intensity is proportional proteins in the sample (Biuret reaction).
CALCULATION
O.D. Sample Total protein = O.D. Standard g/dl: n = 5 g/I: n = 50 12 mmol/l 0.6 mmol/l 30 mmol/l xn
REAGENTS
Reagent 1 Alkalin Potassium-Sodium tartrate reagent Sodium hydroxide Potassium iodide Reagent 2 Colouring Coopersulfate (Nocive) reagent Reagent 3 Standard Bovine albumine
LINEARITY
This method is linear up to 15 g/dl (150 g/I)
REFERENCE VALUES
Newborn 5.2-9.1 g/dl 52-91 g/I 5.4-8.7 g/dl 54-87 g/I 6.7-8.7 g/dl 67-87g/I
0.6 mol/l
Children 5 g/dl 50 g/I Adults

Working Reagent Mix 3 ml of reagent R2 in 1 container R1: Ref 20161 Mix 5 ml of reagent R2 in 1 container R1: Ref 20162 Warning IRRITANT Working reagent is stable : 6 months at +2 - 8C
BIBLIOGRAPHY
Gornall et al. J. Biol-Chem 177, 751 (1949). Henry R.J. Annal. Chem. 92, 1491 (1957) Peter T.J. Clin. Chem.14, 1147 (1968). T.E. Weichselbaum: Am. J. Clin. Pathol. 16. Sect.1040 (1946).
SAMPLES
Serum or heparinized plasma
PROCEDURE
Wavelength : Temperature : Cuvette : Read against blank reagent blank reagent Sample R3: Standard l Working Reagent 1 ml 20l 1 ml 1 ml Standard Sample 20l 546 nm +20 - 25C 1 cm lightpath
Mix and incubate minutes at +20 - 25C Measure the extinction against blank reagent The colour of reaction is stable for 30 minutes
35
FT An 24
Oct 2006
Biomaghreb
PRESENTATION
Ref. 20131(240 Tests) R1 : 2 x120 ml R2 : 2 vials (lyophilized) R3 : 1 x 4 ml Ref. 20132(120 Tests) R1 : 4 X 30 ml R2 : 4 vials (lyophilized) R3 : 1 x 3 ml Rf. 20138 (600Tests) R1 : 5 x 120 ml R2 : 5 vials (lyophilized) R3 : 2 x 5 ml
TRIGLYCERIDES
Enzymatic Colorimetric test (GPO-PAP)
PRINCIPE The triglycrides are enzymatically hydrolyzed to glycrol according to the following reactions : Lipoprotein Lipase Triglycerides Glycerol + Fatty acids Glycerolkinase, Mg++ Glycerol + ATP Glycerol-3-phophate + ADP
Standard Sample Working reagent
Blank _ _ 1 ml
Standard 10 l _ 1 ml
Sample _ 10 l 1 ml
Mix, incubate 5 min. at 37C or 10 min. at 25C. The colour is stable for 30 minutes. CALCULATION O.D. Sample Triglycerides Conc. = O.D. standard mg/dl : n = 200 g/l : n = 2 mmol/l n = 2.28 LINEARITY This method is linear up to 1000 mg/dl (11,4 mmol/l - 10 g/l). If the triglycerides concentration is greater than 1000 mg/dl dilute the sample 1/10 with saline solution and repeat the determination and multiply the result by 10. REFERENCE VALUES Women 40-140 mg/dl 0.40 - 1.40 g/l 0.46 - 1.60 mmol/l 60 - 165 mg/dl 0.60 - 1.65 g/l 0.68 - 1.88 mmol/l xn
3. Glycerol Phosphate Oxidase Glycerol 3-P + 02 Dihydroxyacetone-P +H2O2 Peroxydase H202 + 4-aminophenazone + p-chlorophenol H20 + Quinonimine REAGENTS Reagent 1 Buffer soiLtion
1 Pipes buffer pH 7.2 p-Chlorophenol
50mmol/l 2 mmol/l
Reagent 2 Enzymes
Lipoproteine lipase Glycerolkinase Glycerol-3-P-oxidase Peroxidase 4-Aminophenazone ATP Glycerol
150 000 U/l 800 U/l 4000 U/l 440 U/l 0.7 mmol/l 0.3 mmol/l 200 mg/dl 2 9/1 2.28 mmol/l
Reagent 3 Standard
Men
PREPARATION AND STABILITY Dissolve the contents of one bottle R2, to the contents of one bottle buffer reagent R1. This working-reagent is stable : 4 weeks at 2 - 8C 1 week at 20 - 25C SAMPLES Serum, plasma collected on heparin. PROCEDURE Wavelength : Temperature: Cuvette: Read against blank 505 nm (490-550) 37C (25C) 1 cm lightpath
NOTES The triglycerides in the sample material are stable for 3 days at 2-8C. BIBLIOGRAPHY Fossati, P., Principe, L. Clin. Chem. 28,2077 (1982) Young, D., Pestaner, L. Clin. Chem. 21,5 (1975)
FT An 25
Juin 2008
36
Biomaghreb
PRESENTATION
Ref. 20141, (200 Tests) R1 : 2 x 100ml R2 : 2 vials (Iyophilized) R3 : 1 x 4ml R4 : 2 x 10ml (10xconc.) Ref. 20146, (500 Tests) R1 : 1 x 500ml R2 : 1 vials (Iyophilized) R3 : 1 x 5ml R4 : 1 x 50ml (l0xconc.) Ref. 20148 (1000 Tests) R1 : 2 x 500ml R2 : 2 vials (Iyophilized) R3 : 2 x 5ml R4 : 2 x 50ml (l0xconc.)
Enzymatic colorimetric method (Berthelot modified method)

Blank Standard Sample Reagent A 1 ml Standard 10 l 1ml Sample 10 l 1 ml
UREA
PRINCIPLE
Enzymatic determination of urea to the following reaction : Urea + H20 -> C02 + 2NH3 urease Salicylate and hypochlorite in the reagent react with the ammonium ions to form 2.2 dicarboxy-indophenol. The quantity of this green compound is proportional to the urea concentration.
Mix, incubate 5 min at 37C or 10 min at 20-25C. Then add Reagent4 1 ml 1 ml 1 ml
REAGENTS
Reagent 1 Reagent 2 Buffer Phosphate pH 6,7 EDTA Sodium salicylate Sodium nitroprussiate Urease Standard Urea 60 mmol/l 2 mmol/l 60 mmol/l 32 mmol/l 30000,u/l 50 mg/dl 0,5 9/l 8,325 mmol/l 40 mmol/l 150 mmol/l
Mix, incubate 5 min at 37C or 10 min at 20-25C Then read OD against blank. Coloration is stable 2 hours in dark.
CALCULATION
O.D.Sample Urea = O.D. standard mg/dl : n = 50 g/ l : n = 0.50 mmol/l : n = 8.325 xn
Reagent 3
Reagent 4 (10xconc)
Sodium hypochlorite Sodium hydroxide
LINEARITY
Up to 400 mg/dl (4 g/l - 66,6 mmo/l). In urine up to 100g/l.

Reagent 4 is concentrated. Add 90 ml of distilled water Rf 20141 : 450 ml of distilleed water Ref : 20 146 or Ref : 20 148 Dissolve a vial of R2 in a vial of R1 = reagent A This working solution (Reagent A) is stable: 6 months at 2-8C 2 weeks at 20-25C
REFERENCES VALUES
Serum-Plasma 15-40 mg/dl 0,15-0,40 g/l 2,49-6,66 mmol/l 20-35 g/24h
Urine
SAMPLES
Serum, heparinized plasma. Urine diluted 1 to 50 in distilled water.
BIBILOGRAPHY
Balleter, W.G., Bushman, C.S., Tidwell, P.W., Anal. Chim. 33,59 Berthelot, M.P.E., Repert Chim. Appl. 284 (1859) Mac Key, E.M., Rackeyll, J. Clin. Invest. 4, 295 (1927)
PROCEDURE
Wavelength: Temperature: Cuvette: Read against reagent blank. 590 nm (578 Hg) 25 - 30 - 37C 1 cm light path
37
FT An 26
Juin 2008
Biomaghreb
PRESENTATION
Ref. 20143 (120 Tests) R1 : 4 x 50ml R2 : 4 vials (Iyophilized) R3 : 1x4 ml Ref. 20147 (450 Tests) R1 : 9x50 ml R2 : 9 vials (Iyophilized) R3 : 2x5ml
UREA
Kinetic Test U.V
Pipette into cuvette :
PRINCIPLE
Urea undergs the following enzymatic resction: Working reagent Urease Urea + H20 ---------------------> C02 + 2 NH3 GLDH Standard R3
GLDH 2NH4+ + 2-oxoglutarate + 2 NADH ---------> 2-L- Glutamate + 2 NAD+ +2H20
Standard 1 ml
Sample 1 mI
Incubate at choosen Temperature (25-30-37c) 10 l ----10 l
Sample
The decrease in absorbance due to NADH in unit time is proportional to the urea concentration. GLDH: Glutamate Dehydrogenase
Mix and read OD1 after 20 sec and read OD2 after 80 sec.
CALCULATION
REAGENTS
Reagent 1 buffer solution Reagent 2 Enzymes Tris buffer pH8.0 80 mmol/l
OD Sample
Urea = ------------------ x n OD Standard mg/dl: n = 50 g/l: n = 0,50 mmol/l: n = 8.325
Urease GLDH NADH 2- glutarate
> 10 000 U/L 16 000 U/l 0,30 mmol/l 15 mmol/l
LINEARITY
This method is linear up to 200 mg/dl (33.25 mmol/l 2g/l). If the urea concentration is greater than 2 g/l in Serum or plasma, dilute the sample 1/2 With Saline Solution and repeat assay (result x 2).
Reagent 3 Standard
Standard Urea
50 mg/dl 0,50 g/l 8,325 mmol/l
REFERENCE VALUES PREPARATION AND STABILITY

Reconstitute one vial of reagent 2. With the appropriate volume of buffer/reagent 1. The working reagent is stable 3 weeks at 2-8C 5 days at 20-25C. Serum-Plasma 15 - 40 mg/dl 0,15 - 0,40 g/l 2,49 - 7,47 mmol/l 20 - 35 g/24h
Urine
SAMPLES
Serum, plasma collected on heparin.. Urine diluted 1/100 with distilled water.
NOTES
Do not use anticoagulants containing fluoride or ammonium ions.
PROCEDURE
340nm Wavenlength : 25- 30-37C Temperature : 1 cm lightpath Cuvette : Zero adjustement : air or distilled water.
BIBLIOGRAPHY
Chaney, AL. Clin. Chem 8, 130 (1962) Fawcett, JK. J. Clin. Path. 13, 15 (1960)
FT An 27
Oct 2006
38
Biomaghreb
PRESENTATION
Ref. 20091 (375 tests) R1: 3 x 125ml R2 : 3 vials (Iyophilized) R3 : 1 x 6ml Ref. 20092 (120 Tests) R1 : 4 x 30ml R2 : 4 vials (Iyophilized) R3 : 1 x 4ml Ref. 20095 (240 Tests) R1 : 2 x 120ml R2 : 2 vials(Iyophilized) R3 : 1 x 5ml
URIC ACID
Enzymatic Colorimetric test (Uricase- PAP)
CALCULATION
OD. Sample Uric acid Conc. =----------------------x n OD. standard mg/dl : n=6 mg/l : n = 60 mol/l : n = 357 Multiply result by 10 Urine :
PRINCIPLE Uric acid is oxidized by uricase to allantone and hydrogen peroxide, according to the following equation. Uricase Uric acid + O2 +2H2O ----------> Allantone + CO2 + H2O2 2H2O2 +4 Aminophenazone Peroxydase +2-4 Dichloro-phenol-sulfonate ------------>Quinonemine+4H2O
REAGENTS
Reagent 1 buffer solution Reagent2 vial of enzymes Reagent3 Standard Phosphate buffer pH 7.4 2-4 DHBS Uricase Peroxidase 4-Aminophenazone Uric Acid 50 mmol/l 4 mmol/l 70 U/l 660 U/l 1 mmol/l 6 mg/dl 60 mg/l 357 mol/l
LINEARITY
This method is linear up to 25 mg/dl (1487,5 mol/l - 250 mg/l). If the uric acid concentration is greater than 25 mg/dl in the serum or plasma, dilute the sample 1/2 with saline solution and repeat the determination. Multiply the result by 2.
REFERENCES VALUES
Serum or plasma Women 2.5 - 6.0 mg/dl 25-60 mg/l 148-357 mol/l 3.4 - 7.0 mg/dl 34-70 mg/l 200 - 416 mol/l 250 - 750 mg/24h

- Reconstitute one vial reagent 2 with the appropriate volume of buffer / reagent 1 - This working reagent is stable : 3 weeks at 2-8C 7 days at 20-25C
Men
Urine
SAMPLES
Serum, plasma collected on heparin. Urine diluted 1 to 10 in distilled water.
NOTES
If urine sample is turbid, warm up to about 60C for 10 mn. to dissolve uric acid.
PROCEDURE
Wavelength : Temperature : Cuvette : Blank Standard Sample Working reagent 1 ml 510 nm (490-550) 37C (20-25C) 1 cm lightpath Standard Sample 20 l 1 ml 20 l 1 ml
BIBLIOGRAPHY
Barham and Trinder Analyst 97, 142 (1972) Fossati and Principe Clin. Chem. 28, 227 (1980)
Mix, incubate 5 min. at 37C or 10 min. at 20-25C. The colour is stable for 30 minutes.
39
FT An 28
Oct 2006
Biomaghreb
HEMOSTASIS HEMATOLOGY
Page
APTT
Ref. 30021 6 x 3 ml
42
CALCIUM CHLORIDE
Ref. 30051 5 x 20 ml
42
AUTO FIBRI 1
Ref. 30033 10 x 4 ml
43
AUTO FIBRI 2
Ref. 30034 10 x 4ml
44
CALCIUM THROMBOPLASTIN
Ref. 30011 Ref. 30012 12 x 4 ml 8 x 4 ml
45
FIBRINOGEN
Ref. 30031 Ref. 30032 10 x 2 ml 10 x 4 ml
46
HEMOGLOBIN
Ref. 30041 Ref. 30042 Ref. 30043 50 ml 100 ml 4 x 125 ml
47
41
Biomaghreb
PRESENTATION
Ref. 30021 (180 Tests) 6 Vials of Cephalin-Kaolin (Iyophilized) Auxiliary Reagent Ref. 30051 (1000 Tests) 5 Vials of 20 mis CaCI2 0.025M Ready to use
APTT
Activated Prothrombin time
PRINCIPLE
Cephalin, extract of cerebral lipids use platelet factors as substitude, than explore the intrinsic coagulation pathway. Kaolin is used as factor activation of the contact phase.
RESULTS
Note the clothing time of the patient's plasma and that of the reference normal plasma. Normal values: 30 to 40 seconds Any prolongation of the patient time more than 8 seconds compared with that of the Control need more investigation. In preoperatory investigation, a normal cephalin Kaolin time seems to prevent excessive bleeding. In monitoring anticoagulant therapy by heparin, the patient's CEPHALIN-KAOLIN time is usually 1,5 to 2 times that of the plasma control time.
REAGENTS
Cephalin Kaolin : Dissolue the contents of one vial with 3 ml distilled water. Allow the reconstituted material to stand at room temperature for 30 minutes. Then Swirl gently in order to obtain a homogenous suspension. CaCI 2 : 0.025 M. Read use solution (Code 30051)
STABILITY
Freeze - dried : Reconstituted : 3 years at + 4 C 4 weeks at + 4C Do not freeze
AUXILIARY REAGENT
SAMPLE COLLECTION
Blood (9 vol) is collected in 3,2 % (0.109 M) trisodium citrate anticoagulant (1 vol). Use sample collection tubes made of plastic or siliconized glass. Centrifugation : 20 minutes at 4000 tr/mn Perform the test within 4 hours of specimen collection. Simultaneously collect 2 tubes of healthy donors and perform the test in same conditions. PROCEDURE Realise test in duplicate In a glass test tube at 37 C, add
CALCIUM CHLORIDE
0,025 M
Ready to use
Test sample (control or patient's) Cephalin Kaolin well resuspended Mix, incubate at 37 C for exactly Starting a stop-watch, add 0,025 M
CaCI2 prewarmed at 37 C Mix. Note the clotting time (Sec)
0,1 ml 0,1 ml 3 min
0,1 ml
FT An 29
Juin 2008
42
Biomaghreb
PRESENTATION
Ref. 30033, kit for 400 tests R1 : 10 x 4 ml (Iyophilized) R2 : 1 x 150 ml
AUTO FIBRI 1
Determination of fibrinogen for automated system using electro magnetic detection
In a test tube add : 200 l Dilute plasma Incubate 2 mn at 37C Reagent R1 (incubate at 37C) 200 l Slmultameously, strat a stop watch and note the clot ting time.
PRINCIPLE
Adapatation of the determination of fibrinogen, according to the Clauss method, to detection with automated system. When diluted plasma is exposed to an excess of thrombin, the log of the clotting time bears a linear relationship to the log of the fibrinogen concentration collection.
REAGENTS
Reagent 1 Thrombin Reagent 2 diluent Bovin thrombin Veronal buffer pH 7.35
Automated Technic
At time of test dilute the plasma in Reagent 2 : Fibrinemia Low normal elevated Dilutions 1: 5 1:10 1:20 Plasma (ml) 0.1 0.1 0.1 Reagent2 0.4 0.9 1.9
Stability and storage : at 2-8C until the expiration date given on the pakaging.
SAMPLES
Plasma - Collect blood by clean venipuncture in 0.11 mol/l trisodium citrate observing the correct blood to anticoagulant ratio. (9 vol. - 1 vol.) - Centrifuge 10 minutes to 2.500 g or allow to sediment. - Perform the determination within 6 hours of specimen In a disposable cup. Add. Diluted plasma or capillary blood Incubate for 2 minutes at 37C Place the cups in the measuring chamber Reagent 1, not incubated, well-mixed Read the clotting time and find the mean. 100 l 200 l

Reconstitute 1 vial with vol of distilled water indicated on Reagent 1. Stability after reconstitution : - 8 hours at 20-25C - 48 hours at 2-8C - 1 month at - 20C Congele by fractionized portions in plastic containers Bring the working reagent to 20-25C before the test but do not in plastic container incubate. Mix thoroughly before use
RESULTS
For each clotting time, read off the value of fibrinogen by referring to the correspondence table specific to each lot provided in the kit. Correction for liquid anticoagulant: - If the hematocrit is normal, add 20 % to the result obtained from the table (i. e., multiply the result read on the table by 1.20). - If the hematocrit is abnormal, multiply the result obtained from the table by the correction factor (C). C = 10 - 9 H ( H represents the hematocrit values) 9-9H
PROCEDURE
Manual Technic Dilute to 1/10 plasma with Reagent R2 to obtain a clotting time between 8-25 sec (fibrinemia : 1,5 - 4g/l). If the clotting time is less than 8 sec, dilute plasma to 1/20 or 1/30 than multiply results respectely by 2 or 3. If the clotting time is more than 25 sec, dilute plasma to 1/5 or 1/2 than divise results respectively by 2 or 5. Perform test in duplicate.
REFERENCES VALUES
2.5 to 4 g/l
BIBLIOGRAPHY
Andrew M. et al. Blood 70, 165 (1987). Claussa. Acta Haemat, 17,137 (1957) Caen J. et al. L'expansion scientifique. Paris (1975).
43
FT An 30
Oct 2006
Biomaghreb
PRESENTATION
Ref. 30034, kit for 200 to 400 Tests R1 : 10 x 4 m, (Iyophilized) R2 : 1 x 150 ml
AUTO FIBRI 2
Determination of fibrinogen for automated system using optic detection
In a test tube. Add. Dilute plasma Incubate 2 mn at 37c Reagent R1 (incutiate at 37) 200 l 200 l
PRINCIPLE
Adapatation of the determination, of fibrinogen. according to the Clauss method, le detection with automater system. When diluted plasma is exposed to an excess of thrombin, the log of the clotting time bears a linea relationship to the log of the fibrinogen concentration.
REAGENTS
Reagent 1 Thrombin Reagent 2 diluent Simultaneously. strat a stop watch and note the clotting time. Bovin thrombin + Kaolin 120 NIH U/ml Veronal buffer pH 7.35 Fibrinemia Dilutions 1:10 1:20 1:40 Plasma (ml) 1 0.1 0.05 0.05 Reagent 2 (ml) 0.9 0.95 1.95
Automated technic
At time of test dilute the plasma in .
SAMPLES
- Collect blood by clean venipuncture in 0.11 mol/l trisodium citrate observing the correct blood to anticoagulant ratio. (9vol. - 1 vol.) - Centrifuge 10 minutes at 2.500 9 or allow to sediment. - Perform the determination within 6 hours of specimen cllection.
Low normal elevated
In a disposable up. Add. Diluted plasma or capillary blood 0.2 ml

Reconstitute 1 vial with vol. of distilled water indicated on Reagent 1. Stability after reconstitution : - 8 hours at 20-25C - 48 hours at 2-8C 1 month at - 20C Bring the working reagent to 20-25C before the test but do not in plastic container incubate. Mix thoroughly before use PROCEDURE Manual Technic Dilute to 1/10 plasma with Reagent R2 to obtain a clottin time between 8-25 sec (fibrinemia : 1,5 - 4g/l). If the clotting time is less thaut 8 sec. dilute plasma to 1/20 or 1/30 than multiply results respectely by 2 or 3. If the clotting time is more than 25 sec, dilute plasma to 1/5 or 1/2 than divise results respectively by 2 or 5.
Incubate for 2 minutes at 37C Place the cups in the measuring chamber Reagent 1, not incubated, well-mixed 0.1ml Read the clotting time and find the mean.
RESULTS
For each clotting time, read off the value of fibrinoger by referring to the correspondence tabie specific to each lot provided in the kit. Correction for liquid anticoagulant - If the hematocrit is normal, add 20 % to the resul obtained from the table (i. e., multiply the result read on the table by 1.20). - If the hematocrit is abnormal, multiply the result obtained from the table by the correction factor (C). 10 - 9 H C= (H represents the hematocrit values 9 - 9 H 10 - 9 H
Perform test in duplicate
REFERENCES VALUES
2.5 to 4 9/1
BIBLIOGRAPHY
ndrew M. et al. Blood 70, 165 (19B7). Claussa. Acta Haemat, 17,137 (1957) CAEN J. et al. L'expansion scientifique. Paris (1975). FT An 31 Oct 2006 44
Biomaghreb
PRESENTATION
Ref. 30011, kit for 240 tests 12 x 4 ml (Iyophilised) Ref. 30012, kit for 160 Tests 8 x 4 ml (Iyophilised)
CALCIUM THROMBOPLASTIN
Dtermination of the prothrombin Time PT
Reconstituded : 30 days at 2 - 8 C 8 hours at 37C Do not freeze the reagent.
PRINCIPLE Quick time is a global test to measure activiy of extrinsic coagulations factors (Factors II, V, Vll and X). The principle consist to measure the clotting time of the patients plasma and to compare it with that of a normal human plasma in presence of excess cacium and tissular thromboplastin. SAMPLE COLLECTION Blood (9 vol) is collecded in trisodium citrate anticoagulant (1 vol). Centrifuge blood sample for 10 minutes at 2500 g or allow to sediment. Test must be done within 6 hours after sample collection. REAGENT Dissolve a vial of the Calcium Thromboplastin reagent in a vial of solvant. Swirl gently in order to obtain a homogeneous suspension. CALIBRATION Consist to converte quick time expressed in seconds to prothrombine rate expressed in % in comparaison with normal value (Thivolle Line). A Pooled nomal human plasma is diluted to 1/2, 1/3, 1/4, 1/8 in Michaelis buffer solution.
PROCEDURE Perform test in duplicate Bring thromboplastin 30 min to 37C before use. The patients samples are used undiluted In a glass test tube at 37 C add : Plasma normal or patients Incubate at 37 C for approximately 2 mn Starting a stop-watch. add thromboplastin prewarmed at 37C Mix. Note the clottingtime Note the clotting time using thivolle line. Converte times obtained to prothrombine rate. In monitoring oral anticoagulatant therapy by AVK, results can be expressed by INR (International Normalized Ratio) defined af follow: INR = 200 l 100 l
Dilutions 1/1 Plasma (ml) Tampon (ml) Activity % 100
1/2 0,5 0,5 50
1/3 0,5 1 33
1/4 0,5 1,5 25
1/8 0,5 3,5 12,5
Times Patient Times Controle
ISI
Construct the Thivolle line on linear graph paper by plotting the reciprocal of each standard dilution on the X - axis, and the corresponding clotting time (in seconds) on the Y - axis. Use the special graph paper provided in the kit. STORAGE AND STABILITY Freeze-dried : The reagents are stable until the expiry date indicated, when stored at 2-8c.
ISI : International sensitivity index is calibrated against the World health organization thromboplastine Reference preparation. ISC is determined and referred to the correspondance table for each lot and provided in the kit. NORMAL VALUES Normal activity: 70 to 100 % A PT of more than 100 % has no pathologied significance. Therapeutic range of antivitamin K treatment between 20 to 30 %
45
FT An 32
Juin 2008
Biomaghreb
PRESENTATION
Ref. 30031, kit for 100-200 Tests R1 :10 x 2 ml (Iyoph) R2 :1 x 100 ml Ref. 30032 kit for 200-400 lests R1: 10x4 ml (Iyoph) R2 :1 x 150 ml Ref 30035 kit for 250-500 Tests R1 : 5x10 ml( Iyoph) R2 :1 x 150 ml
FIBRINOGEN
2- Technic with fibrometer
In a disposable cup of fibrometer at 37C. Add Diluted plasma or capillary blood 0.2 ml Incubate for 2 minutes at 37C Reagent 1, not incubated, well-mixed 0.2 ml Read the clotting time
PRINCIPLE
Determination of fibrinogen, according to the Clauss method. When diluted plasma is exposed to an excess of thrombin, the log of the clotting time bears a linear relationship to the log of the fibrinogen concentration.
REAGENTS
Reagent 1 Thrombin Reagent 2 diluent Bovin thrombin Veronal buffer pH 7.35
3- Automated technic : electro magnetic detection

At time of test dilute the plasma in Reagent 2:
Fibrinemia Dilutions Plasma (ml) Reagent 2
SAMPLES
- Collect blood by clean venipuncture in 0.11 M trisodium citrate observing the correct blood to anticoagulant ratio. ( 9vol. -1 vol. ) - Centrifuge 10 minutes at 2.500 g or allow to sediment. - Perform the determination within 6 hours of specimen collection.
Low normal elevated
1: 5 1:10 1:20
0,1 0,1 0,1

100 l
0,4 0,9 1.9
In a disposable cup.Add. Diluted plasma or capillary blood Incubate for 1 minutes at 37C Place the cups in the measuring chamber Reagent1, not incubated, well-mixed Read the clotting time and find the mean.
50 l
4- Automated technic : Optic detection

At time of test dilute the plasma in :
Fibrinemia Dilutions Plasma (ml) Reagent 2

Reconstitute 1 vial with vol of distilled water indicated on Reagent 1. Stability after reconstitution : - 8 hours at 20-25C - 48 hours at 2-8C - 1 month at - 20C Keep the reconstituted reagent R1 in its original vial befor the test to 20-25C but do not in plastic contain incubate. Congele by fractionized portions.
Low normal elevated
1: 10 1: 20 1: 40
0.1 0.05 0.05

100l
0.9 0.95 1.95
In a disposable up.Add. Diluted plasma or capillary blood Incubate for 1 minutes at 37C Place the cups in the measuring chamber Reagent1, not incubated, well-mixed Read the clotting time and find the mean.
50 l
PROCEDURE
Dilute to 1/10 plasma with reagent R2 to obtain a clotting time between 8 and 25 sec (fibrinemia 1,54 g/l). If the clotting time is less than 8 sec, dilute plasma to 1/20 or 1/30 than multiply results respectively by 2 or 5 respectively. If the clotting time is more than 25 sec, dilute plasma to 1/5 or 1/2 than divise results respectively by 2 or 5.
N.B : To improve the optical dtection of the coogulation, it is necessary to use Kaolin in 10% (reconstitue the lyophilisat by 2 ml of water distilled for the reference 30031 and add 10l of kaolin 10% in this vial : it is the working reagent) The Kaolin is supplies in the demand.
RESULTS
For each cloiting time, read off the value of fibrinogen by referring to the correspondence table specific to each lot or provided in the kit. Correction for liquid anticoagulant: - If the hematocrit is normal, add 20 % to the result obtained from the table (i. e., multiply the result read on the table by 1.20). - If the hematocrit is abnormal, multiply the result obtained from the table by the correction factor (C) .
1- Manual technic
In a glasse test-tube add Dilute plasma Incubate at 37C for 2 mn. Starting a top watch add. add. Reagent R1 prewurmed at 37C 200 l 200 l
C=
10 - 9 H 9-9H
( H represents the hematocrit values)
Dip the hook into the liquid in the center of the tube and move the hook up and down regularly until a thin filament of fibrin a pears. Note the clothing time. Start stop watch and note the clotting time. FT An 33 February 2009 46
REFERENCES VALUES
2.5 to 4 g/1
BIBLIOGRAPHY
CLAUSS A. Acta haematol 17, 137 (1957) CAEN J. et al. Blood 70, 165 (1987).
Biomaghreb
PRESENTATION
Ref. 30041, kit for 500 tests R1 : 1 x 50 ml Ref. 30043, kit for 5000 Tests R1 : 4 x 125ml Ref. 30042, kit for 1000 Tests R1 : 1 x 100ml
HEMOGLOBIN
Color method
CALCULATION PRINCIPE
Heamoglobin is oxidesed to cyanmethemoglobin with DRABKIN Reagent. Hemoglobine conc. g/l = OD Sample x 376
REAGENTS
Reagent 1 Drabkin Reagent 50 fold concentrad Potassium Ferricyanur Potassium cyanur Phosphate monopotatium Sterox 30 mmol/l 38 mmol/l 50 mmol/l 25 g/l
REFERENCES VALUES
(Biochemists Handbook 1961) Newborn : Children (1 year) : Children (10 years) : Man : Women : 195 50 g/l (12 mmol/l) 112 g/l (6.95 mmol/l) 129 g/l (8 mmol/l) 160 20 g/l (9.9 mmol/l) 140 20 g/l (8.7 mmol/l)
Toxic Reagent: use automatic pipettor. Store at 20-25
SAMPLE
Blood collected on whole EDTA. PROCEDURE Work reagent : Drabkin Reagent R1 Distilled water 1 volume 49 volume
BIBLIOGRAPHIE
Brit J. Haemat. 13, 71. (1967) Drabkin D.L et al.- J.Biol. Chem. 98, 719. (1932), Zijistra N.C - Clin. Chim. Acta, 5,719. (1960)
Stability
1 month at 20 - 25 C (do not refrigerate) Wavelength Blank 540 nm (546 Hg) work reagent Sample Sample work solution Mix and Measure the OD. Coloration stability 1 hour 20 I 5 ml
(avoid to expose Reaction on strong light)
47
FT An 34
Oct 2006
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IMMUNOHEMATOLOGY
Page
ANTI- A MONOCLONAL ANTIBODY

Ref. 30111 Ref. 30112 10 ml 10 x 10 ml
50
ANTI- AB MONOCLONAL ANTIBODY

Ref. 30131 Ref. 30132 10 ml 10 x 10 ml
51
ANTI - B MONOCLONAL ANTIBODY

Ref. 30121 Rf. 30122 10 ml 10 x 10 ml
52
(IgM + IgG) ANTI - D MONOCLONAL ANTIBODY 53 Ref. 30141 Ref. 30143 10 ml 10 x 10 ml
POLYVALENTE ANTIGLOBULIN Ref. 30181 Ref. 30182

49
54 10 ml 5 ml
Biomaghreb
PRESENTATION
Ref. 30111, Drop bottle for 100 at 400 tests Ref. 30112, Drop bottle for 1000 at 4000 tests
REAGENT :
Mouse monoclonal antibody.
ANTI - A MONOCLONAL ANTIBODY

Tube test method :
- Wash 3 times the red blood cells in isotonic saline solution. - In a tube mix 2 drops of reagent and 2 drops of 5% cell suspension in isotonic saline. - Centrifuge for1 minute at 500g. - Resuspend the cells gently . - Read visually
STORAGE :
The reagent is stable at + 4C until the expiry date stated on the kit box. Sodium azide is used as preservative at 1%o Reagent only for in vitro diagnostic use.
SAMPLE :
Preferably take blood on dry tubes with anticoagulant, dissociate well the clot we have to conserve sample at +4C which can not be examined immediately. (No haemolyse must be observed).
PARTICULAR ATTENTION :
The anti-A reagent reacts with A3 red cells.
METHODS :
On not heated plate (at + 20C) * No washed Total Blood. - Place 2 drops of reagent beside 1 drop of total blood (not washed). - With adapted agitator mix on centrifuge spiral movement to form a circular reaction with 2 cm in diameter. - Rock several times the plate to homogenize the mixture. - Read visually for 3 minutes. * No washed Red blood cells in suspension at 10 % in isotonic saline : - Place 2 drops of reagent beside 2 drops of red blood suspension (not washed)at 10% in isotonic saline - With adapted agitator mix on centrifuge spiral move ment to form a circular reaction with 2 cm in diameter. - Rock several times the plate to homogenize the mixture. - Read visually for 3 minutes. Plate test method : - Wash 3 times the red blood cells in isotonic saline solution . - Add into a well 25l of reagent and 25 p1 of 5% cell suspension in isotonic saline. - To homogenize the mixture slightly agitate the plate with adapted agitator . - Incubate for 10 minute at + 20C.. - Centrifuge for 1 minute at 200 - 300 g. - Agitate the plate until resuspending of the negative control. (25 l of reagent replaced by 25 l of isotonic saline solution ). - Read visually.
FT An 35
Oct 2006
50
Biomaghreb
PRESENTATION
ANTI -AB MONOCLONAL ANTIBODY

Tube test method :
REAGENT
STORAGE
The reagent is stable at + 4C until the expiry date stated on the kit box. Sodium azide is used as preservative at 1 Reagent only for in vitro diagnostic use.
SAMPLE
preferably take blood on dry tubes with anticoagulant, dissociate weli the clot we have to conserve sample at +4C which cannot be examined immediately. (No haemolyse must be observed).
The anti-A reagent reacts with Ax red cells.
METHODS :
On not heated plate (at + 20C) * No washed Total Blood. - Place 2 drops of reagent beside 1 drop of total blood (not washed). - With adapted agitator mix on centrifuge spiral movement to form a circular reaction with 2 cm in diameter. - Rock several times the plate to homogenize the mixture. - Read visually for 3 minutes. * No washed Red blood cells in suspension at 10 % in isotonic saline: - Place 2 drops of reagent beside 2 drops of red blood suspension (not washed) at 10% in isotonic saline - With adapted agitator mix on centrifuge spiral movement to form a circular reaction with 2 cm in diameter - Rock several times the plate to homogenize the mixture. - Read visually for 3 minutes.
Plate test method :

- Wash 3 times the red blood cells in isotonic saline solution. - Add into a well 25 l of reagent and 25 l of 5% cell suspension in isotonic saline. - To homogenize the mixture slightly agitate the plate with adapted agitator . - Incubate for 10 minute at + 20C.. - Gentrifuge for 1 minute at 200 - 300g. - Agitate the plate until resuspending of the negative - control. (25 l of reagent replaced by 25 l of isotonic saline solution). - Read visually .
51
FT An 36
Oct 2006
Biomaghreb
PRESENTATION
REAGENT :
ANTI - B MONOCLONAL ANTIBODY

Tube test method :
STORAGE:
SAMPLE :
Preferably take blood on ciry tubes with anticoagulant, dissociate well the clot we have to conserve sample at +4C which cannot be examined immediately. (No haemolyse must be observed).
The anti-A reagent reacts with B3 red cells.
METHODS :
On not heated plate (at + 20C) * No washed Total Blood. - Place 2 drops of reagent beside 1 drop of total blood (not washed). - With adapted agitator mix on centrifuge spiral movement to form a circular reaction with 2 cm in diameter. - Rock several times the plate to homogenize the mixture. - Read visually for3 minutes. * No washed Red b/ood cells in suspension at 10 % in isotonic saline: - Place 2 drops of reagent beside 2 drops of red blood suspension (not washed) at 10% in isotonic saline - With adapted agitator mix on centrifuge spiral movement to form a circular reaction with 2 cm in diameter. - Rock several times the plate to homogenize the mixture. - Read visually for 3 minutes. Plate test method : - Wash 3 times the red blood cells in isotonic saline solution . - Add into a well 25 l of reagent and 25 pl of 5% cell suspension in isotonic saline. - To homogenize the mixture slightly agitate the plate With adapted agitator . - Incubate for 10 minute at + 20C.. - Centrifuge for 1 minute at 200 - 300g. - Agitate the plate until resuspending of the negative control. (25 l of reagent replaced by 25 l of isotonic saline solution). - Read visually .
FT An 37
Oct 2006
52
Biomaghreb
PRESENTATION
REAGENT :
Human monoclonal antibody in albumin medium.
(IgM + IgG ) ANTI - D MONOCLONAL ANTI BODY in albumin medium

polyvalent antiglobulin. (follow the antiglobulin procedure). - Centrifuge at 500 g for 1 minute. Resuspend the cells gently. - Read visually.
STORAGE:
SAMPLE :
Take blood on tube with anticoagulant. we have to conserve sample at +4C which can not be examined immediately. (No haemolyse must be observed).
PARTICULAR ATTENTION
In indirect coombs the albuminal monoclonal anti-D reagent reacts with low D-samples. For any sample it should be to use a negative albuminal control. The no agglutination with the negative control confirms the specificity of a positive reactivity of the anti - D reagent
METHODS :
Heating slide test (at + 40C) or Rhesuscope test. - place 1 drop of reagent beside 1 drop of total blood (not washed). - With adapted agitator mix on centrifuge spiral movement to form a circular reaction with 2 cm in diameter. - Rock several times the plate to homogenize the mixture. - Read visually for 3 times. Plate test method at 37 C. - Wash 3 times the red cells in isotonic saiine solution. - Make a 5% cell suspension in isotonic saline solution. - Add into a well 25 l of reagent and 25 l of cell suspension. - To homogenize the mixture, slightly agitate the plate with adapted agitator. - Incubate for 10 minutes at 37C (to avoid evaporation cover the plate with lid ) - Centrifuge for 1 minute at 200 - 300 g. - Agitate the plate until resuspending of the negative control. - Read visually. Indirect coombs test : - Wash 3 times the red cells in isotonic saline solution. - Make a 3 - 5% cell suspension in isotonic saline solution or in low isotonic strength solution(LISS). - Into test tube mix 2 drops of reagent and 2 drops of ceil suspension. - Incubate at 37C for 45 minutes (if isotonic saline solution is used) or 15 minutes (if low isotonic strength solution is used). - Wash 3 times in isotonic saline solution and dry the pellet of last washing before adding 1 drop of IgG or
PRECAUTION
Humain origin product were found negative for HBs Ag and for antibodies against HCV, HIV-1 and HIV-2 by approved test methods. However,since no test method can offer complete assurance that infectious agent are absent, this product should be handled observing the same safety precautions employed when handling any potentilly infectious material.
53
FT An 38
Oct 2006
Biomaghreb
PRESENTATION
Ref. 30181, Drop bottle for 100 tests Ref. 30182, Drop bottle for 50 tests
POLYVALENTE ANTIGLOBULIN
REAGENT FOR COOMBS TEST
REAGENT
Animal serum origin
STORAGE
At +4C until expiry date stated indicated on the kit box. sodium Azide at 1 is used as preservative. Only for in vitro diagnostic use. A positive reaction is performed only if negative controls and autotogous controls are negative. A negative reaction is performed only if positive controls is positive (confirming the reactivity of the antiglobulin).
METHODS
Standard indirect Coombs test Prepare a 5 % cell suspension in isotonic NaCI solution of red blood cells preliminary washed 3 times in isotonic NaCI solution. In test tube, mix 2 drops of serum (or reagent) and 2 drops of 5% cell suspension. Incubate for 45 mn at 37C. Wash 3 times in isotonic NaCI solution, dry the pellet after the last washing. Add one drop of polyvalente antiglobulin. Centrifuge at 500 9 for 1 mn. Resuspend the cells gently and read macroscopically for agglutination or microscopically after displaying on lamina. Indirect coombs test using Liss Buffer (Low isotonic saline solution). Use the technique describe above but cells are 3% resuspended in LiSS Buffer. Incubate for 15 mn at 37C. Direct Coombs test Use same method but suppress the sensibilization of red cells step.
INTERPRETATION
Serums controls must be used as fellow. - Indirect Coombs test : Autologous Control : Red celis and patient serum. - Direct Coombs test : Negative control in saline : The antiglobulin is replaced by isotonic NaCI solution. Negative AB Control serum : The antiglobulin is replaced by the AB serum. Positive control : Sensibilized red cells with diluted or low nown antibody.
FT An 39
Oct 2006
54
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SEROLOGY AND QUICK TESTS

Page
ASO LATEX
Ref. 40037 Ref. 40023 50 Tests 100 Tests
56
CRP LATEX
57
RHEUMATOID FACTORS LATEX

58
STREPTOLYSINE- O
Rf. 40021 Rf. 40022 6 x 3 doses 10 x 3 doses
59
TPHA
Ref. 40011 Ref. 40012 100 test 300 test
61
UCG 25
Ref. 40032 Ref. 40033 Ref. 40034 20 bandeletts 100 bandeletts 40 bandeletts
62
55
Biomaghreb
PRESENTATION
Ref. 40037kit for 50 tests Ref. 40023kit for 100 tests
ASO LATEX
Test for the determination of anti-streptolysin O in serum samples
SEMI-QUANTITATIVE TECHNIQUE :
Prepare serial two-fold dilutions in physiological saline and Observe the presence or absence of agglutination. The approximate anti- SLO level in serum sample can be calculated by the following formula. ASO Titre lU/ml = Highest dilution with positive reaction X reagent sensitivity (200 lU/ml). NORMAL VALUE Up to 200 lU/ml.
SUMMARY
Anti-Streptolysin O antibodies are produced in those infections promoted by beta-hemolytice Streptococci, due to the presence the Streptolysin O (SLO) liberated from the bacteria. Information on the extent and degree of the infection can be obtained from the measure of the serum level antibodies.
PRINCIPLE
ASO-Latex reagent is a suspension of polystyrene particles sensitized streptolysin 0. When the reagent is faced against, the serum with anti-SLO antibodies, and antigen - antibody reaction takes place being easily visualized because of the latex agglutination.
INTERPRETATION
Streptococcol A infections disease can be complicated by acute Rheumatoid arthitis, acute glomenula nephritis. Essential biological diagnostic is immunologic-as no bacteria was found in late streptococci infection. The rate's anti-streptolynine O antibodies is more than 200 lUtml in 80% of streptococci infections. A repeated test is recommended 15 days later as 200 lU/ml represents the pathological limit detection.
REAG ENTS
Aso latex Reagent Positive control Negative control Stirrer, Test slide Latex suspension Dropper-bottle ready to use Dropper-bottle ready to use
BIBLIOGRAPHY
Alouf, J.E. and Raymond, M.: Biochimie, 56 (1973). Bach, G. Wiatr, R.A., Anderson, T.O. and Cheatle, E.: Amer.J. Clin. Path., 52 (196g). Halbert, S.P.: Ann. N.Y. Acad. Sci. 103 (1963). Klein, G.C., Manual of Clin. Immunoligy, Amer. Soc. Microbiol p.264 (1976). Klein, G.C., Baker C.N. and Jones, W.L. Appl. Microbiol., 21, 999 (1979). Schmidt, k., Mueller-Echardt, Ch. And Beckmann, A.: Rheumatol, 29 (1970)
STORAGE AND STABILITY

The component of the kit, when stored at 2 - 8C, will remain stable until the expiration date stated on the label. Do not freeze. Discard those control in which, in spite of containing sodium azide, a microbial growth is apparent.
SAMPLES
Fresh serum or stored at 2 - 8C for no longer than 48h. It is necessary to freeze the sample when the assay is to be carried out after the period of time. Discard contaminated, lipemic and highly haemolized sera, as well as plasma interfere with the assay.
PROCEDURE
1-Bring reagents and serum sample to room temperature. 2-Place one drop of undiluted serum out to a slide black area. 3-Mix well the latex reagent and add one drop over the serum drop. 4-Mix with the aid of stirrer both drops and tilt the slide. 5-Observe the presence or absence of agglutination within a period not longer than 3 minutes under a strong light. 6-Occasional agglutinations produced after 4 min have no diagnostic significance.
RESULTS
Latex agglutination will mean that the serum anti-SLO level is higher than 200 lU/ml.
FT An 40
Oct 2006
56
Biomaghreb
PRESENTATION
Ref. 40034 Ref. 40043 50 tests 100 tests
CRP DIRECT LATEX REAGENT

Test for the determination of C-Reactive protein in serum
SEMI QUANTITATIVE TECHNIQUE
Prepare serial two-fold dilutions in physiological saline and observe the presence or absence of agglutination. The approximate C-Reactive protein leveMn serum sample can be calculated by the following formula : CRP mg/l = Highest dilution with positive reaction X reagent sensitivity (6 mg/l). NORMAL VALUE Up to 6-8 mg/l
SUMMARY
C-Reactive protein is a globulin, which increases in inflammatory processes, in the acute phase of different diseases and after surgical operations. The main diagnostic value of its determination is the detection of infiammatory processes having a rheumatic origin: acute articular rheumatism or chronic polyarthrytis in its acute phase.
PRINCIPLE
CRP-latex reagent is a suspension of polystyrene particles sensitized with anti-human C-Reactive protein. When the reagent is faced against the serum with C-Reactive protein, an antigen antibody reaction takes place being easily visualized because of the latex agglutination.
LIMITATION OF THE PROCEDURE

Occasional agglutinations produced after 4 min have no dia gnostic significance. Lipemic and highly haemolized sera, as well as plasma inter fere with the assay. Rheumatoid factors in the sample may also agglutinate the latex reagent. Prozone phenomena appear at 80 g/ml.
REAGENTS
CRP Latex reagent Positive control Negative control Glass Slide'Stirrer CRP suspension Dropper-bottle, ready to use. Dropper-bottle, ready to use.

The components of the kit, when stored, at 2 - 8C will remain stabie until the expiration date stated on the lable. Do not freeze. Discard those control in which, in spite of containing sodium azide, a microbial growth is apparent.
BIBLIOGRAPHY
Kidmari, C.O. (1972). Scand. J. Clin. Invest., 29, 407-411. Deyo, R.A., Pope, R.M., Perselin, R.H. (1980). J. Rheumatol., 7, 279-287.
SAMPLE
Fresh serum or stored at 2 - 8 C for no longer than 48h. It is necessary to freeze the sample when the assay is to be carried out after pehod of time. Discard contamined or hemolyzed sera .
PROCEDURE
1- Bring reagents and serum sample to room temperature. 2- Place one drop of undiluted serum onto a slide black area. 3- Mix well the latex reagent and add one drop over the serum. 4- Mix with the aid of stirrer both drops and tilt the slide. 5- Observe the presence or absence of agglutination within a period not longer than 3 minutes.
INTERPRETATION
Latex agglutination will mean that the serum C-Reactive protein level is higher than 6,0 mg/L. In patients with high serum concentrations continued monitoring of the CRP levels can give a good indication of patient response to the therapy during inflammtory disorders.
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FT An 41
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PRESENTATION
Ref. 400 36 Ref. 400 24 50 Tests 100 Tests
PRINCIPLE
RHEUMATOID FACTORS LATEX is a rapid latex agglutination test kit for the detection of Rheumatoid Factor (RF) in human serum. The RF latex particles are coated with specially purified human gamma globulin. When the latex suspension is mixed with serum containing elevated RF levels on a slide, clear agglutination is seen within 2 minutes.
RHEUMATOID FACTORS LATEX

Latex serology test for detection of Rheumatoid Factor
RESULTS
Examine the test slide under a strong light source after 2 minutes. A positive result is indicated by the obvious agglutination pattern of the latex, in a clear solution. A negative result is indicated by no change in the latex suspension on the test slide then indicate a RF concentration below 6 lU/ml.
REAGENTS
Latex Reagent The Latex suspension should be well shaken before use; Dropper-bottle Ready to use Dropper-bottle Ready to use
SEMI QUANTITATIVE TEST

Prepare a serial dilution of patient's serum in diluant buffer (1/20,1/40,1/80 and 1/160). Repeat the test procedure for each dilution as described above. The serum RF concentration can then be calculated approximately by multiplying the dilution factor by the detection limit, to give the number of lU/ml concentration e.g. if the agglutination titre appears at 116 the approximate serum RF concentration is 6 x 6= 36 lU/ml.
Positive Control Serum
Negative Control Serum
Stirrers Test slide RHEUMATOID FACTORS LATEX Kits should be stored at 2-8c.
INTERPRETATION
In suspected clinical Rheumatoid arthritis, the Rhumatoid factors are detected in 80% patient's serum, seropositive forms are serious in compared with seronegative forms. In positive test, confirm results by Waaler Rose reaction. Rhumatoid factors are also found at 4 % of patients with lupus, hepatitis, syphilis and various other clinical conditions. These positive results give very low titres compared to Rheumatoid arthritis.
SAMPLES
Fresh or freeze serum within 1 month. Do not use haemolysed, contaminated or lipaemic sera for testing.
PROCEDURE
Allow test reagents and sera to come to room temperature.
BIBLIOGRAPHY
Ball, J. and Lawrence, J.S.: Ann. Rheum. Dis., 22, 311 (1963). Jones, W.L. and Wiggins, G.L.: Amer, J.CLin. Path., 60, 603 11973) Singer, J.M. and Plotz, G.M.: Am. J. Med. 21, 888, (1956). Singer, J.M. and Plotz, G?M.: JAMA, 168, 180 (1958). Waaler, S.G., et al. P.: Bull. Wld. Hlth, Org.42,311 (1970).
OUALITATITIVE TEST
Transfer one drop of patient's serum and one drop of each control to separate circles on the test slide. Shake the latex reagent, then, using the dropper provided, add one drop of suspension to each circle. Mix the drops using a disposable stirrer ensuring coverage of the test circle with the mixture. Gently and evenly, rock and rotate the test slide for 2 minutes whilst examining the test slide for agglutination.
FT An 42
Oct 2006
58
Biomaghreb
PRESENTATION
Ref. 400 21 kit for 16 Tests Ref. 400 22 kit for 27 Tests
STREPTOLYSIN - O
Determination of anti streptolyin - O
MACROMETHOD
Samples without hemolysis and inactivated by heating at 56C for 30 mn. If the inactivation exceed 24 hours, it's recommanded to heat again the serum at 56C for 10 mn/
PRINCIPLE
A,C,G groups of haemolytic streptococcus secreted an enzyme : Streptolysin- O. wich present an haemolytic activity in its reduced form. This streptolysin-O causes antibody formation revealed by a neutralization reaction of its enzymatic haemolytic acitivity towards rabitt red blood cells.
REAGENTS
- Lyophylisat of titrated streptolysin - Streptolysin Buffer : 10 x concentrated, to dilute before use : 1 volume of streptolysin Buffer 9 volumes of distilled water If cristalization, put the streptolysin buffer at 37C before use. -1 % Rabitt cell suspension. Carefully washed. Human O red cells can be used. - Titrated streptolysin reconstitution : Add exactely 8 ml of streptolysine buffer 1 x to the vial without lose Iyophilisat material (use a needie to introduce aire befor opening the vial). Close the vial and shake gently. The streptolysin is in its activated form.
PROCEDURE
Primary dilutions Into two tubes, A and B, introduced 2 ml of Streptolysin buffer 1 X. In Tube A add 0.25 ml of serum to test (dilu tion to t : 5) mix and pipette 1 ml of tube A into tube B (dilution to 1: 15). Secondary dilutions For each serum, prepare tubes in duplicate : n tube 1,2,4,6,8,10 -> for primary dilution n tube 3,5,7,9 -> for tube B primary dilution n tubes 11,12 - Erythrocyte and streptolysin controls. dilute serums as descnded in table 1 by geometrical progress (in ml volume repartition).
STORAGE
Lyophilised streptolysin : 3 years at +4C Reconstitued streptolysin : 5 hours
Tubes n
Dilutions Streptolysine BufferIX tampon Dilution A (srum 1/5) Dilution B (srum 1/15)
1
1/5
2
1/10 0,25
4
1/20 0,25 0,25
6
1/40 0,25 0,25
8
1/80 0,25 0,25
10
1/160 0,25 0,25
3
1/15
5
1/30 0,25
7
1/60 0,25
9
1/120 0,25
11
Eryth 0,5
12
Strepto 0,25
0,25
0,25
0,25 0,25
0,25
0,25 0,25
0,25 0,25 0,25
ReconstitutedStreptolysin 0,25 Erythocytes 1% suspension
0,25 0,25 0,25 0,25 0,25 0,25 Mix and leave 10 mn at room temperature 0,25 0,25 0,25 0,25 0,25 0,25
0,25
0,25
0,25
0,25
0,25
0,25
leave 10 mn at room temperature then 20mn at 37 in water bath. Centrifuge at 1500 rpm for 3 mn ASLO amount according to the last tube without hemolysis
50
100
200
400
800
1600
150
300
600
1200
non lysis
Total lysis
Usual values are less than 200 ASU samples wiht value equal or greater than 200 ASU are considered pathological.
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FT An 43
Oct 2006
Biomaghreb
MICROMETHODE Make dilutions of serums to test :
- 1: 5 : Dilution : 0,1 ml of serum + 0,4 mol of streptolysine buffer 1 x - 1:15 : dilutions : 0,1 ml of serum + 1,4 mol of streptolysine buffer 1 x Distribute into the wells of microplaque titration
Colonns Wells N srum dilution Streptolysine Buffer 1 x 1 : 5 dilution serum 1 : 15 dilution serum Mix Reconstituted Streptolysine
-->
Colonns 5 6 1 2 3 4 5 Controls
1 1/5
1/10 1/20 1/40 1/80 1/60 1/15 1/30 1/60 1/120 1/240 Erytroc. Strepto 0,05 0,05 0,05 0,05 0,05 0,05 0,05 0,05 0,05 0,10 0,05
0,05 0,05 0,05

0,05- > 0,05- > 0,05- > 0,05- >
0,05
--> 0,05-> 0,05 - > 0,05 - > -
0,05
0,05
0,05
0,05
0,05
0,05 0,05
0,05
0,05
0,05 0,05
0,05
Mix well and leave 20 mn at room temperature

Erytrocyte 1 % Suspension (in left to right) 0,05 0,05 0,05 0,05 0,05 0,05 0,05 0,05 0,05 0,05 0,05 0,05 0,05
Leave 10 mn at room temperature than 20 mn at 37C Centrifuge the plate for 2 mn at 500 g or read hemolysis by lift up and observating under the plate or leave sediment at 18- 25 C
ASLO amount according to the test well wthout hemolysis
50
100
200
400
800
1600 150
300
600
1200 2400
No lysis
Total lysis
Usual values are less than 200 ASU. Samples with value equal or greater than 200 ASU are considered pathological.
60
Biomaghreb
PRESENTATION
Ref. 40011, kit for 1 00 Tests Ref. 40012, kit for 300 Tests
TPHA
Haemagglutination Test for the Serodiagnostic of Syphilis
READING and INTERPRETATION
Examine macroscopically the selling patterns of the cells after 45 mn incubation. Results are stable until 1 day Readings are scored and reported according to the following criteria. Smooth mat of cells covering entire botton of well Smooth mat of cells covering part of the well bottom Smoot!h mat of cells sunounded by red cricle surronded by a smaller red circle Define compact button of cells. 2+ 1+ -. Smooth mat of cells covering less area and 3+ 4+
PRINCIPLE
Avian erythrocytes sensilized with treponema palladium antigen will agglutunate in the presence of serum antitreopnema antibodies to give characteristic patterns micro titration plate. No specific reactions are defined by erythrocytess control (without senzitisation).
REAGENTS
-Test Cells : Stabilized avian erythocyte suspension senzitized with T. pallidum antigen 1 vial of 7,5 ml - Control cells : Stabilized avian cells suspension not sensitized 1 vial of 7,5 ml - Diluant Buffer : 1 vial of 20 ml - Positive controi serum : Pre-diluted at 1/20 1 vial of 1 ml - Negative control serum : pre-diluted at 1/10 1 vial of 1 ml - V.U. Well microtitration plates : 3 plates
SAMPLES
The sea should be stored at 2-8C. They can be freezed and defreezed only once. decomplementation is not necessary. The sera containing particles in suspension should be centrifuged.
PROCEDURE
Bring the test reagents and samples to room temperature
QUALITATIVE TEST
Well 1: Dilute serum 1/20 with Diluent Buffer : 10 I serum + 190 l Buffer.. - Mix and distribute 25 I of this dilution into well 2 et 3. - Well 2 added 75 l of control cells - Well 3 added 75 ,l of test cells - Mix by thorongly tapping all four sides of the plate. - Cover the plate an incubate at room temperature for 45 minutes. Keep the plate away from vibration, near and direct sunlight. Do not dilute positive and negative control seum they are ready to use with test and Control Cells.
The Negative Control must show a non-agglutinated eattern with both Test and Control Cells. positive Control must show anglutination with Test Cells but not with the Control Cells. Sera showing agglutination with the Control Cells contain non-specific agglutinins and shouid be retested after absorphon. To absorb sera, mix 100 ,ul of serum with 400 ,l of Control Cells in a small test tube. Mix-and incubate at room temperature (15-30C) for 1 hour. Centrifuge at 1000 r.p.m. (600 x g) for 5 minutes and relest a 1/4 diluhon of the supermatant in Diluent (Dilution 1/20).
TEST INTEREST
The TPHA has early positive results, apparing only a few days after the positive FTA. Positive reactions indicate late or ancient svphilis infectious, date infectious is characterized by tftre increase confirmed 10-20 days later. Evolutive syphilis is indicated by the 2 fold increase of dilution titre.
QUANTITATIVE TEST
Pipette into adjacent wells of the microtitation plate:
Well Buffer Serum 1/20 Well 1 (l) Test cells l Dilution 1/N 25 75 80 25 75 160 75 320 75 640 75 75 75 75 1280 2560 5120 10240 A B 25 C 25 25 D 25 25 E 25 25 F 25 25 G 25 25 H 25 25
BIBLIOGRAPHY
Rathlev T. Brit. J. Vener. Dis, 1957, 43 381. Tomizawa T., Kasamatsu S., Japori. Med. Sci. Biol. 1966, 19.30 S. Paris-Hamelin A., Vaisman A., Fustec Ibarboure S. Pharm -, Biol., 1982, XVI, 179.
Mix cover the plate and incubate at room temps for 45 minutes. Keep the plate away for vibration,heat and direct sunlight.
61
FT An 44
Juin 2008
Biomaghreb
PRESENTATION
Ref. 40032 40033 40034 20 Tests 100 Tests 40 Tests
UCG 25
Dipsticks
The UCG 25 Dipstick Pregnancy test, is a rapid qualitative, on step assay for the detection of human chorionic gronadotropin in serum and urine.
PROCEDURE
- Remove the required number of reaction strips dipsticks from dessicated container and close it firmly again. - Dip the reaction strip into the tube containing the sample. Do not dip over the red mark. - Hold the dipstick for 10 sec. - Remove the strip from the specimen and place it on the plane surface. - Read the results after 5 to 10 mn.
PRINCIPLE
This method uses the CICA method (Concured Immuno chromatography Assay). A monoclonal anti HCG antibody bound to color particules is fixed on a membrane itself fixed on the dipstick (solide phase). As the test sample flows through the absorbent portion of dipsticks : 1-In the presence of the labelled antibody dye conjugate binds to HCG forming an antibody-antigen complex. This complex binds to the anti-HCG antibody in the posivite reaction zone and procedure a pink-rose color band. The reaction mixture continues flowing through the absorbent portion of dipstick past the positive reaction zone and control zone. Unbound conjugate binds to the reagents in the control zone, producing a pink-rose color band, demonstrating that the rengents are functioning correctly. 2- In the absence of HCG, there is no line in the positive reaction zone. Only the control zone presents a pink rose color band demonstrating that the reagent are functioning correctly.
READING TEST-RESULTS
Negative test
One colored band closest to the top (control band) will appear to show that the test is complete.
Positive test
In addition to the control band, a clearly distinguishable band will also appear closest to the bottom ~ndicating presence of HCti in sample.
Inconclusive test
If there is no distinct color band visible both in the top and the bottom area of the reaction dipstick. It is recommended in this case that the test be repeated or a fresh specimen be obtained and tested.
SENSITIVITY
UCG 25 detects in less than 5mn, levels of HCG as low as 25 mlU/ml (OMS 3st standard). Specimens containing high levels of HCG (200 000 m lU/ml) when tested consistently gave positive results.
CROSS REACTIVITY
Following concentration of homologous hormons are found to have no interferences with Biomaghreb's UCG25; h TS H 100 mlUlml h LH 500 m iU/ml h FSH 1000 m I Ulml

The UCG 25 kit is to be stored at room temperature (4c to 30C). Do not freeze the tests kit. Only for in vitro diagnostic use.
BIBLIOGRAPHY
Braunstein, G.D., Rasor J., Adier, D., Danzer H, and Wade, M.E. Am. J. Obstet. Gynecol., 126, 678-681 (1976). Braunstein, G.D., waitukaitis, J.L., Carbone, P.P., and Ross, G.T., Ann. Inter. Med., 78, 39-45. (1973). Engvall, E., Methods in Enzymology, 70, 419-439 (1980). Lenton, E.A. Neal, L.M., and Sulaiman, R., Fertility and Sterility, 37, 773-778 (1982). Morgan, F.J., Canfield, R.E., Vaitukeitis, J.L., and Tompson, R.J., Jackson, A.P., and langlois, N., Clin Chem., 32, 476-481 (1986). Rasor, J.L., and Braunstein, G.D. Obstet. GYnecol, 50, 553558 (1977). Ross, G.T. Endocrinologv,94, 1601-1606 (1974). Kohier, G. and Milstein, C., Nature 256, 495-497 (1975),
SPECIMEN COLLECTION AND PREPARATION

1- Serum
- Avoid hemolysis - If the test is to be run within 48 hours after collection, the specimen should be stored in the refrigerator (+4C+8C). - If testing is detayed more than 48 hours, the specimen should be frozen. - The specimen must be completely thawed, througly mixed and brought to room temperature priorto testing.
2- Urine
The first morning urine specimen is prefered since it contains the highest concentration of HCG. - Collect the urine specimen in a clean container without preservatives. - The specimen should be refrigerated (4+8c) for up 2 days. In such case bring the specimen to room temperature prior to testing. If testing is delayed more than 24 hours, the specimen should be frozen. The frozen specimen must be completely thawed and brought to room temperature prior to testing.
FT An 45
Oct 2008
62
Biomaghreb
CONTROL 5 minutes
SERUM OR PLASMA
Dip the ship for 10 sec and place it on the plane surface
NEGATIVE
POSITIVE
63

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Biomaghreb

PRICE QUALITY and SERVICES

20101 20103 20161 20062 30011 40032 20141 20142 20091

INORGANIC PHOSPHORUS UV METHOD

TOTAL IRON - BINDING CAPACITY

0.62 mmol/l 69 mmol/l

and multiply the result by 2.

PREPARATION AND STABILITY

40 mmol/l 1 mg/l 17,9 mol/l

PREPARATION AND STABILITY

Anyway each laboratory should assign their own reference values.

PREPARATION AND STABILITY

Blank Standard Sample Working reagen 1 ml

Urine 680 nm 1 cm light path 20-25C against reagent blank

PREPARATION OF REAGENTS AND STABILITY

TOTAL IRON-BINDING CAPACITY

Note : Use preferably plastic throwaway material.

Basic magnesium carbonate

PREPARATION OF WORKING SOLUTIONS AND STABILITY

BIOCHEMICAL ENZYMES Page

AMINO TRANSFERASES COLORIMETRIC METHOD

23 20011 20012 200123 200125 20 x 3 10 x 10 9 x 50 15 x 10 ml ml ml ml

Kinetic Test. IFCC Without Pyridoxal phosphate

3 ml (25,30 a 37C) 300 I

Equilibrate at choosed temperature. Sample Mix and wait 1 mn. 100 l

10 mmol/I 25C 30C 37C

Reagent 2 GPT substrate

95 mmol/l 200 mmol/l 2 mmol/l

Reagent 3 Colour reagent Reagent 4 standard

Mix. Let stand for 20 minutes at room temperature.

Mix. Let stand for 20 minutes at room temperature. NaOH 0.4 N 10 10 10 10 10

Mix. Wait 5 minutes, Measure. FT An 09 Oct 2006 18

30C Up to 22 U/I Up to 26 U/I

37C Up to 31 U/l Up to 38 U/I

+ NADPH+H The catalyte activity of Ck is determined by the measurement of NADPH formation.

2- Manual procedure : Substrate start

PREPARATION AND STABILITY

Anyway each laboratory should assign their own references values.

PREPARATION AND STABILITY

PREPARATION AND STABILITY

CALIBRATOR FOR TOTAL AND DIRECT BILIRUBIN

TOTAL AND DIRECT BILIRUBIN

TOTAL BILIRUBIN TOTAL PROTEINS TRIGLYCERIDES

UREA (COLORIMETRIC METHOD)

UREA (KINETIC METHOD)

REFERENCE VALUES STORAGE AND STABILITY

Sample Standard Reagent 1

CALIBRATEUR BILIRUBINE TOTALE ET DIRECTE

Apres reconstitution a lobscurite - 2 jours a 20-25c - 4 jours a 2-8c - 6 semaines a - 20c

Pipette into tests tubes Blank 1 ml Standard 10 l 1ml Sample 10 l 1 ml

Standard Sample Working reagent

Reagent 2 vial of enzymes

PREPARATION AND STABILITY

Increased risk above :

Read against blank reagent, standard and sample.

Cholesterol CHOD-PAP Assay

HDL Cholesterol < 0,35 g/l < 0,9 mmol/l)

492 nm (490-510) 25, 30 ou 37C 1 cm lightpath

Standard Blank Standard R4 Sample Reagent R2 Working solution 50 l 1 ml 1 ml Reac 50 l

Sample Blank Reac

505 nm (490-550) (20, 25, 37C) 1 cm lightpath

TOTAL AND DIRECT BILIRUBIN

CALCULATION (TB and DB)