FATIMA JINNAH CHEST AND GENERAL HOSPITAL QUETTA

PROVINCIAL PUBLIC HEALTH LABORATORY

MEASLES ELISA IgM

Date: ___________

Purpose:___________________ Performed by : __________

Buffer Preparation : Per 100 ml = 5ml w/sl x + 95ml D/W = 100 ml

Washing Solution for 1 Reaction : 300 x 5 = 1500 x 5 = 7500ul

Preparation Date:____________ Lot#___________

1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H
TEST PROCEDURE: IgM

1. Add 25ul of Vircell IgM sorbent only in test wells

2. Add 5ul sample and 75ul of serum diluents (Total 105ul)

For Control

 100ul serum diluents + 5ul positive control

 100ul serum diluents + 5ul negative control
 100ul serum diluents + 5ul cut off control
 (Over with sheet and incubate at 370C for 45 minutes)
 Wash 5times + add 100ul IgM conjugate (incubate 370C for 30min)
 After that wash 5 times and add 100ul substrate solution
(incubate 370C for 20min)
 Add 50ul stopping solution
 Read the absorbance 450/620nm
 FATIMA JINNAH CHEST AND GENERAL HOSPITAL QUETTA
PROVINCIAL PUBLIC HEALTH LABORATORY
MEASLES ELISA IgG

Date: ___________

Purpose:___________________ Performed by : __________

Buffer Preparation : Per 100 ml = 5ml w/sl x + 95ml D/W = 100 ml

Washing Solution for 1 Reaction : 300 x 5 = 1500 x 5 = 7500ul

Preparation Date:____________ Lot#___________

1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H
Test PROCEDURE: IgG
1. Add serum diluents 100ul in each well.
2. Add 5ul serum in test well
3. Add 5ul of positive control + 5ul of negative control+ 5ul of cut off
control (in each well total 105ul)
4. Incubate at 37 0C for 45min
5. Wash 5times and add 100ul of IgG conjugate.
6. Incubate at 37 0C for 30min
7. Wash 5 times
8. Add 100ul substrate (Incubate for 20 min at 370C)
9. Add 50 ul stopping solution.
10. Read with spectrophotometer at 450/620nm