Serology o Primary infection

- Ab and Ag reaction in VITRO.  Localized infection

- Purpose:  Appearance of painless, hard
o Unable to culture infection agents chancre
o Confirmation of etiologic agent  Soft chancre, painful
identification  H.ducreyi
 Using serological method  LAB TEST: Dark filed microscopy
o Diagnosis of immunologically-related (presence of coiled organism with
disorders corkscrew motility.
 Ex: Autoimmune disorders  NO SEROLOGICAL TEST
o Determine the immune status  EARLY INFECTION: No antibody
 Ex: immunity to Rubella, Hepa B. production
etc.  10-90 days (21 days average)
Serologic Test o Secondary infection (vry infectious stage)
- Types:  Characterized by generalized rash
o Detect unknown Ag in specimen by using and lesions.
known Ab  Apperance of Condylomata lata
o Detect unknown Ab in specimen by using (flat-wartz like lesions)
known Ag.  LAB TEST: Dark field microscopy
and Serology test (production of
Bacterial Serological Test Ab in the body)
I.Syphilis/Great Pox/ Evil Pox/ Italian Pox, French Pox, o Latent stage
Spanish Pox  Not infectious
- By Treponema pallidum subspp. Pallidum  Generally after 2nd infection
- Christopher Colombus- spreads the Syphilis.  Asymptomatic
- Corkscrew motility, filamentous, helically unicellular  Indication of the infection is
bulb positive serological test.
- Subsspp. speciation  (+) Serologic test but no sign and
o T.p.sbsp. pallidum- Syphilis symptom.
o T.p.sbsp pertenue- Yaws  LAB TEST: Serologic test, no dark
o T.p.sbsp endemicum- non-venereal field microscopy? No lesions.
endemic syphilis/ Bejel
o T.p.sbsp. carateum- Pinta o Tertiary infection
- Indistinguishable to each other:  Presence of gummas (destructive
o Morphologic lesions)
o Serological test  Gummas are dead
- Differentiation is based on: treponema, and not
o Based on clinical and epidemiological infective
characteristic.  MANIFESTATION:
- CHARACTERISTICS:  Gammatous syphilis
o Too thin to be seen with light microscopy in  Cardiovascular disease
specimens stained with Gram stain or  Neurosyphilis
Giemsa stain.  TREATMENT:
o Intra cellular pathogen  1st choice is a heavy
 Cannot be grow in cell for cultures metal- Arsenic
in vitro  2nd choice is a drug-
 Do not survive outside of host Penicillin
o It has TROMPS (treponemal rare outlier - Other manifestation of tertiary infection
membrane proteins o Neurosyphilis
 Delayed immune system of host  Asymptomatic
 A protein  Involvement of CNS and detected
- MOT: only By CSF examination
o Sexual intercourse o Congenital syphilis
o Parenteral exposure  Caused by maternal spirochetemia
o Congenital infection during pregnancy and transplacental transmission of
- CLINICAL MANIFESTATION: microorganism
  Western blot- test of choice  Neutralizes complementary
 MANIFESTATION: properties of cardiolipin
 Hutchinson’s teeth
 Intersitial keratitis o Cholesterol
 Nerve deafness  Provides adsorption centers and
 Other manifestations: increase the effective reaction of
 Fussing around the mouth cardiolipin.
and arms - REPORTING:
 Skeletal lesions o Non-reactive = no clumps
 Perforation of the palates o Weakly reactive= small clumps
and collapsed nasal bones o Reactive= medium to large clumps
(Saddle nose deformity)  Report the last reactive
- FALSE (+) CDRL Results (ShRIMP)
DIAGNOSTIC MARKERS: o SLE
o RF
- Reagin antibodies (non-treponemal marker) o IM
o Screening test o Malaria
o Non-treponemal Ab o Pregnancy
o Non-specific
o Not sure if it’s a Treponema pallidum sbspp. 2. RPR (Rapid Plasma Reagin)
Pallidum
o Cardiolipin - Uses unheated serum
 Antigen used in the test - Result is read MACROSCOPICALLY
 From cow’s heart ak.a Wasserman - PRINCIPLE: Flocculation
antigen or Diphospatidyl glycerol - REAGENT:
 Wasserman Test 1st test used to o Colorless alcoholic solution containing
detect syphilis. cardiolipin
- Anti-treponemal Ab o Lecithin
o Confirmatory test o Charcoal- for visualization
o Specific o Choline chloride- chemical inactivator
o Treponemal Ag- Ag used o Thimerosal- preservative

SEROLOGICAL TEST FOR SYPHILIS ROTATION GAUGE DROPS RING

OF DM
A. Non-treponemal/Non-specific methods NEEDLE

- Used to detect Reagin (an antibody which is against VDRL

the common Ag of T.pallidum
- PRINCIPLE: Complement fixation -Quali 180 rpm 4 18 60 14 mm
o (+) cell lysis mins

1. VDRL (Venereal Disease Research Laboratory Test) -Quanti 180 rpm 4 19 75 14mm
mins
- Uses heated serum and CSF
o Serum is heated for the inactivation of -CSF 180 rpm 8 21/22 100 16 mm
complement activities mins
o Inactivated- 10 mins
o Heated 56°C at 30 minutes RPR 100 rpm 8 20 60/75 18mm
- Results is read MICROSCOPICALLY mins
- PRINCIPLE: Flocculation
- CSF is not heated? No complement activity
- REAGENT:
o Cardiolipin (diphosphatidyl glycerol) B. Specific method/Treponemal method
 Main reagent
o Lecithin - Uses true treponemal Antigen and Antibody
- 2 Sources:
 o Non-pathogenic: - Gram (+) cocci
 Reiter’s strain- Removes cross- - Streptoccal Antigen
reactivity with treponemas o Streptolysin O
o Pathogenic  Most pathogenic
 Nichol’s Strain- fixation  Bacterial toxin released during
- Absorbent: Reiter’s treponemal infection
 Oxygen LABILE
1. FTA-ABS (Flourescent Treponemal Antibody-Absoprtion)  HIGHLY ANTIGENIC
 Hemolytically inactive in oxidized
- Test for dark field microscopy form
- PRINCIPLE: Indirect fluorescence Immunoassay  Neutralized by ASO
- REAGENT: Nichol’s strain dried and fixed on slide o Streptolysin S
 Causing clearing in the agar
2. HATTS (Hemagglutination Treponemal Test for Syphilis - CLINICAL MANIFESTATION:
o Streptoccocus toxic shock syndrome
- PRINCIPLE: Hemagglutination o Pharyngitis
- REAGENT: Glutaraldehyde stabilized turkey RBC o Erysipela: Impetigo: Skin infection
coated with Treponemal antigen o Scarlet fever (Dick’s Test)
 (+) Strawberry tongue
3. MHA-TP (Microhemagglutination Treponemal pallidum  By Erythrogenic toxic
Test)  Exotoxin B (dangerous)

- PRINCIPLE:Hemagglutination Serological Test for Group A. Strep

- REAGENT: Tanned formalin sheep RBC coated with
Treponemal antigen ASO Testing (Macrotechnique of Rantz and Randall)

4. Treponemal pallidum Immobilization Test - PRINCIPLE:: neutralization (no hemolysis)

- Most specific test
- Standard Test
- PRINCIPLE: The antibody produced against
Treponemal pallidum plus complement can
immobilize the live treponemas.
- REAGENT: Live actively motile T.p organism
(extracted from Lesions of infected rabbits
- RESULTS: (+) >50% immobilized treponema
o If <50%? Not considered as + result
o 20-50%- doubtful, repeat the test
o <20- negative
- This test estimates amount of the antibody that is
presence of a constant dose of SO can completely
inhibit hemolysis of a constant given no. of red cells.
- TITER: Reciprocal of Highest dilution demonstrating
no hemolysis.
- Expressed in Todd Units or International Unit
- CONTROL:
o Red cell control tube (+) control
o SO control tube (-) control
- Normal range:
o Children- <125 todd units
o Adults- <166 todd units

Other Strep Test

Rapid Latex Agglutination Test

II. Group A Streptococcal infection - PRINCIPLE: Passive agglutination

- SIGNIFICANT TITER: >200 IU/mL
- Streptococcus pyogenes- pus producing
- Protein M- inhibits Ceb (stops the complement Anti-DNAse B Testing
cascade
- Cocci in chains - Highly specific for group A streptococcal sequelae
- Common in upper respiratory tract - PRINCIPLE: Neutralization
- COMPLICATION:
o Acute glomerulonephritis Streptoenzyme Testing
o Rheumatic fever
- Lancefield sero Group A - Agglutination test for Group A
- B-hemolytic pattern - Excellent test
- Slide agglutination technique - Rickettsial organism
- PRINCIPLE: Agglutination - Gram (-) OBLIGATE intracellular organism
- MOT:
III. Febrile Agglutinins o Athropod bite or contamination with feces
- Antibody demonstrated in microbial diseases that of Ticks
are manifested by high fever - Weil Felix Test
o Typhoid Fever o Non-specific test which detects Antibody
o Typhus Fever against Rickettsial disease.
o Brucellosis o PRINCIPLE: Direct agglutination
o Tularemia o Antibody: Rickettsial organism
o Antigen: Proteus antigen
Interpretations of bacterial agglutinations test  Proteus vuglaris
 OX-19 and OX-2
- A rapid slide test is only for screening a (+) test  Proteus mirabilis
should be confirmed by tube test.  OX-K
- A single (+) test is of no significance unless the titer is
high

TYPHOID FEVER Rickettsial OX-19 OX-2 OX-K

Disease
- Acquried by: Ingestion
- Salmonella typhi Scrub- - - ++++
- Spread by Typhoid Mary Mannon- based on my O.tsutsugamuchi
memory tinapay na may peanut butter yung ginawa
nya dun kumalat. RMSF-R.rickettsi ++++ + -
- Salmonelle Antigen (HKO Virluence)
o H and K Antigen (Vir A) Epidemic
 Thermolabile typhus-
o O Antigen (common) R.prowazecki-
 Thermostable pathogenic to
o H=Hauch- Flagellar human
o K=Kapsel- Capsular
o O= Ohne- Somatic Murine typhus-
o R.mooseri
- LAB DIAGNOSIS
o Culture method Q-fever-Coxiella - - -
 1st two weeks- Blood burnetii
 3rd week- Urine
 4th week- Stool Rickettsial- - - -
o Widal Test R.akari
 For rapid diagnosis of Typhoid
fever LYME DISEASE
 Slide test- principle: Direct
agglutination - Lyme is a place where it 1st isolated
 Tube test- Highest serum dilution - CAUSATIVE AGENT: Borrelia burgdorferi
showing ++ or 50% agglutination - TRANSMITTED VIA: Ixodex spp.
 SIGNIFICANT TITER:80 and above - LAB DIAGNOSIS:
o Immunofluorescence
o Typhidot o EIA
 Detection of specific IgM and IgG o WB- confirmatory test
to Salmonella typhi o PCR
 IgM/G against Outer - CHARACTERISTIC: Bull’s eye rashes
membrane protein
 All steps based on ELISA LEPTOSPIROSIS

TYPHUS FEVER - CAUSATIVE AGENT: Leptospirosis interrogans

- Rodent’s urine 
Development of anti-HAV IgG and
- INFECTIVE STAGES: immunity on recovery
o Septicemic stage o LAB DIAGNOSIS:
 Fever headache, mya  RIA
o Immunological stage  ELISA
 Jaundice  ARKITEK
 Azotemia- ↑ Urea - Hepatitis B
- LAB DIAGNOSIS: o Agent HBV
o Culture method- Fletcher or Stuart Media o Dane Particle (infectious form)
o Microscopy- dark field microscopy  Leads to liver cirrhosis and cancer
o Serological test o PRE VACCINATION SCREENING

HbsAg Anti- Anti- Status Action

HBs Hbc
MYCOPLASMA PNEUMONIAE
+ - + Acute/chronic No immunization
- “Eaton agent” infection
- PAP
- Without cell wall? Requires STEROL - + + Resolved No immunization
- Leading cause of upper respiratory infection world infection
wide
- Fried-egg appearance - + - Previous Follow up
- Walking pneumonia vaccination
- DIAGNOSIS: Complement fixation
o ELISA + - - Acute No immunization
o Cold agglutination test infection
- PRINCIPLE: Clinical samples with M.pneumoniae
infection contains IgM antibodys that target the I - - + Window Active
Antigen of RBC and induce agglutination period immunization
- TRANSPORT MEDIAS:
o TSB with 0.5% Albumin - - - No detectable
o SP4 medium previous
o Viral transport medium exposure to
 Delay in plating? Frozen at -70°C HBV
(for weeks)
- Hepatitis C (NANB)
o Test:
 Surrogate Test- ALT/SGPT and anti-
VIRAL DISEASE HBC
 Serologic Test for anti HCV
Hepatitis Virus Disorders  ELISA
 RIA
o Primary- A B C D E F G  RIBA- HCV specific Ab / anti-HCV
o Secondary- EBV,SLE,IM,CMV,HSV CHON
- Hepatitis A  RT-PCR- HCVRNA persistent HCV
o Philippine setting: Most common Hepatitis infection
is B - Hepatitis D
o Book based: Most common is Hepatitis A. o Anti-HBc IgM differentiates CO-infection to
o Former name: Enterovirus No. 72 super infection
o HEPA.A MARKERS OF INFECTIONS o Test: PCR- confirmatory
 Sheds in stool o CO-INFECTION
 Appearance of IgM anti HAV onset  Severe acute disease
of symptoms  Low risk of chronic infection
 Icterus and Increase liver o SUPER INFECTION
enzyme levels  Usually develop chronic HDV
infection high risk of severe
 chronic liver disease may present
as an acute hepatitis.
- Hepatitis E
o LAB DIAGNOSIS:
 Electromicroscopy
 Indirect ELISA
 RT-PCR
- Hepatitis G
o Blood borne hepatitis
o Causes Syntical Giant Cell hepatitis
o Flavivirus like Hepa. C
o NO SEROLOGICAL TEST
IM (Infectious Mononucleosis)
- Kissing disease or Glandular fever
- AGENT: EBV
- TARGET CELL: B-lymphocytes
- COMMON IN: Adults and adolescence
- Antigens:
o EA- early antigen
 EA diffuse- seen in Nucelus and
Cytoplasm
 EA restricted- seen in cytoplasm
o NA- nuclear antigen
o EBNA- EBV Nuclear Ag
- Heterophile Antibodies
o Refer to Antibodies produces as part of an
immune response to an infection but that
are not related to the causative agent.
- CHARACTERISTICS DIAGNOSTIC PROFILE OF EBV.
STAGE Description
If the patient is
Susceptibility seronegative (lacks of
antibody to VCA)
Ab (IgM) to VCA is
Primary infection present: EBNA is absent,
high or rising titer or Ab
(IgG) to VCA and no
evidence of Ab to EBNA
after at least 4 weeks of
symptoms
If Ab to EBNA and
Reactivation increased antibodies to
EA are present
Ab to VCA and EBNA are
Past infection present
HIV (Human immunodeficiency Virus)
- CLINICAL SYMPTOMS:
o Primary
 2-4 weeks after infections
 Flu-like symptoms “worst flu ever”
 “acute retroviral syndrome” (ARS)
or “primary HIV infection”
o Clinical Latency
 No symptoms
 Asymptomatic HIV/Chronic HIV
 Negative results pero
nakaka hawa pa din
o AIDS
 Opportunistic infection
 CD4 cells <200 cells / mm3
- SCREENING TEST:
o ELISA
o RIA
o Agglutination tests
- CONFIRMATORY
o WB- (+) if gp 41 bands appearance or when
an envelope Ab (gp 41, 120, or gp 160)
appears in combination with another HIV
o BOOK BASED: atleast 2 bands (+) or 3
o SOME LAB: DAPAT 3 BANDS
Notes to remember..
- Testing is often done at 6 weeks, 3 months or 5
months after exposure to find out if a person is
infected with HIV
- The US CDC defines POSITIVE results for HIV as
“repeatedly WB”
o 2 ELISA and 1 WB
DENGUE
- CLINICAL MANIFESTATION:
o Thrombopenia
o Hemoconcentration
o (+) tourniquet test
- LAB DIAGNOSIS:
o Complement fixation test
o Immunoassay
- Hemorrhagic dengue virus
o (+) sa unang sero test, tas pag nag ka
dengue ulit at nag positive, yan ung pwede
maging sakit ng patient.
- Infected by dengue
o Loose blood vessels
o Plasma lalabas, then plasma sasama niya
platelets kaya decrease platelets.
- Treatment: FFP, not platelet concentrate
o Clot blood vessels
- Dehydration may lead into plasma breakage, so
drink more water
- Serological Test
o Rapid NSI Dengue Test
 Use if 1 to 2 days infection
o Rapid Dengue anti-IgM/igG
 Use if 5 days or more
 NSI test may be negative
 (+) IgM- present infection
  (+) IgG- past infection
 (+) IgM/igM- recovery

RUBELLA

- Pink rash
- Rubivirus
- Respiratory secretions
- CLINICAL DIAGNOSIS:
o ELISA- screening test
o PCR- confirmatory test
o Hemagglutinaton
RUBEOLA
- Mumps
- Paramyxoviridae
- Koplik spots
o Negri bodies? Lyssa virus
- DIAGNOSIS:
o Viral culture
o Serology