IMMUNOLOGY

and
SEROLOGY
SEROLOGY OF BACTERIAL INFECTION

Serology of Syphilis
Streptococcal Serology
Febrile Disease Serology
Serology of Syphilis
 Serology of Syphilis
▪ Human syphilis is caused by the spirochete Treponema pallidum.

▪ Antibodies against treponemal antigens and nontreponemal

cardiolipin antigens (Wasser-mann antigens) develop and elicit a
cell-mediated and humoral Immune response.

▪ Transmission of Disease: Sexual Contact

Blood or blood-product transfusion
Woman to her Fetus.
 Serology of Syphilis
FOUR STAGES OF SYPHILIS:

1. Primary (Early) Syphilis

2. Secondary Syphilis
3. Latent Stage of Syphilis
4. Tertiary Syphilis
 Inflammatory lesions (chancres) appear 2 to 8
Primary Syphilis weeks after infection and last for 1 to 5 weeks.
Usually occurs 6 to 8 weeks after chancres
first appear. This stage is characterized by a
Secondary Syphilis generalized rash, and secondary lesions may
develop in the eyes, joints, or central nervous
system (CNS).
Highly contagious and is generally considered
Latent Stage to begin after the second year of infection.
Is characterized by granulomatous lesions
known as gummata. These lesions may
Tertiary Syphilis develop in skin, mucous membranes, joints,
muscles, and bones.
 Inflammatory lesions (chancres) appear 2 to 8
Primary Syphilis weeks after infection and last for 1 to 5 weeks.
Usually occurs 6 to 8 weeks after chancres
first appear. This stage is characterized by a
Secondary Syphilis generalized rash, and secondary lesions may
develop in the eyes, joints, or central nervous
system (CNS).
Highly contagious and is generally considered
Latent Stage to begin after the second year of infection.
Is characterized by granulomatous lesions
known as gummata. These lesions may
Tertiary Syphilis develop in skin, mucous membranes, joints,
muscles, and bones.
 Serology of Syphilis
CONGENITAL SYPHILIS
Syphilis can be transmitted to a fetus after the 18th week of
gestation.

Treatment of the infected mother before the 18th week will

prevent infection; treatment after the 18th week will cure it.
 Serology of Syphilis
Note:

Treatment. Penicillin is the drug of choice, although tetracycline

or erythromycin can also be used.

Primary Stage: A seropositive patient in the primary stage of

disease usually becomes nonreactive approximately 6 months
after treatment.
 Serology of Syphilis
Note:

Secondary Stage: If treatment occurs during the secondary

stage, the patient usually becomes nonreactive within 12 to 18
months after treatment. Patients treated 10 years or more after
infection may always remain seropositive.
 Serology of Syphilis
Tests for Syphilis are based on the detection:

1. NON-TREPONEMAL ANTIBODY DETECTION:

a. Venereal Disease Research Laboratory (VDRL)
b. Rapid Plasma Reagin (RPR) Test
2. TREPONEMAL ANTIBODY DETECTION:
a. Fluorescent Treponemal Antibody Absorption Test (FTA-ABS)
b. Treponema pallidum Immobilization (TPI) Test
c. Antibody Capture Enzyme-linked Immunosorbent Assay (ELISA)
d. Hemagglutination Tests
NON-TREPONEMAL ANTIBODY DETECTION
Venereal Disease Research Laboratory (VDRL) Slide Test:
▪ The VDRL is a qualitative and quantitative agglutination test
using heat-inactivated patient serum.
▪ Inactivation of Serum: 56*C for 30 minutes.
▪ Reactivation of Serum: 56*C for 10 minutes.
▪ CSF can also be used.
▪ Uses a slides with ceramic ring.
▪ Antigen: 0.03% Cardiolipin, 0.9% Cholesterol, 0.21% Lecithin
NON-TREPONEMAL ANTIBODY DETECTION
Venereal Disease Research Laboratory (VDRL) Slide Test:
▪ Antigen Delivery Needles:

Qualitative Serum VDRL: 18 gauge needle without bevel

that will deliver 60 drops of
antigen suspension per mL

Ring Diameter = 14 mm
NON-TREPONEMAL ANTIBODY DETECTION
Venereal Disease Research Laboratory (VDRL) Slide Test:
▪ Antigen Delivery Needles:
Quantitative Serum VDRL: 19 gauge needle without bevel
that will deliver 75 drops of
antigen suspension per mL, 23
gauge needle that with or
without bevel that will deliver
100 drops of saline per mL.
Ring Diameter = 14 mm
NON-TREPONEMAL ANTIBODY DETECTION
Venereal Disease Research Laboratory (VDRL) Slide Test:
▪ Antigen Delivery Needles:
CSF VDRL: 21 or 22 gauge needle that will deliver
100 drops per mL.
Ring Diameter = 16 mm
1.75 mm depth
NON-TREPONEMAL ANTIBODY DETECTION
Rapid Plasma Reagin (RPR):
▪ The RPR is an agglutination test. Modified VDRL Test.
▪ Antigen is similar to the VDRL antigen with the addition of the
following: Charcoal, EDTA, Thimerosal, Choline Chloride
▪ Unheated serum is the specimen of choice, although plasma
may be used.
▪ Uses a plastic cards.
▪ Antigen delivery: 20 gauge
▪ Serum: 100 rpm for 8 minutes
NON-TREPONEMAL ANTIBODY DETECTION
VDRL RPR
Method Flocculation Flocculation
Detects Reagin Reagin
Antigen Cardiolipin Cardiolipin with charcoal
Positive Microscopic Macroscopic
Reaction Clumps Agglutination
Specimen Inactivated Serum Serum
CSF
NON-TREPONEMAL ANTIBODY DETECTION
VDRL RPR
Reactivity during May be neg in primary stage. Same as
disease Titers usually peak during VDRL.
secondary or early late stages.
Titers in late stage, even when
untreated. More rapid decline
with treatment. Becomes
nonreactive in 1–2 yr following
successful treatment.
NON-TREPONEMAL ANTIBODY DETECTION
VDRL RPR
False Positives Biologic false pos with Same as
infectious mononucleosis (IM), VDRL.
infectious hepatitis, malaria,
leprosy, lupus erythematosus,
rheumatoid arthritis, advanced
age, pregnancy. Reactive in
other treponemal infections
such as yaws & pinta.
 TREPONEMAL ANTIBODY DETECTION
Fluorescent Treponemal Antibody Absorption Test
(FTA-ABS):

▪ The FTA-ABS test detects treponemal antibodies by

using a killed suspension of T. pallidum as an
antigen and a fluorescein-conjugated antihuman
globulin reagent.
 TREPONEMAL ANTIBODY DETECTION
Treponema pallidum Immobilization Test:

▪ Standard test to which other treponemal test are evaluated.

▪ Involves mixing of patient serum with live, actively motile T.
pallidum extracted from testicular chancre of a rabbit and
complement.
▪ Test is considered positive if ≥ 50 treponemes are
immobilized.
 TREPONEMAL ANTIBODY DETECTION
Treponema pallidum Immobilization Test:

▪ Interpretation:

If 50% or more = Positive

20% to 50% = Doubtful Result
If 20% or less = Negative
 TREPONEMAL ANTIBODY DETECTION
Hemagglutination:

▪ Reagent: RBC’s Sensitized with Nichol’s Strain

▪ Hemmaglutination Treponemal Test for Syphilis (HATTS)

▪ T. pallidum Hemagglutination Assay (TPHA)
▪ Microhemagglutination Assay for Antibodies to T. pallidum
(MHA-TP)
Interpretation of Syphilis Test Results
Streptococcal Serology
 Streptococcal Serology
▪ Streptococcus pyogenes is a gram (+) coccus responsible for a
human infections, some of which can have serious sequelae.

▪ The M protein is the major virulence factor for S. pyogenes.

▪ Bacterial Toxins:
1. Streptolysin O (SLO)
2. Streptolysin S
 Streptococcal Serology
Bacterial Toxins
Streptolysin O (SLO) ▪ Is an oxygen-labile enzyme that
causes hemolysis by binding to
cholesterol in the RBC membrane.
▪ It is antigenic, and the presence of
antibodies to SLO is an indicator of
recent streptococcal infection.
 Streptococcal Serology
Bacterial Toxins
Streptolysin S ▪ Is a non-antigenic, oxygen-stable
enzyme.
▪ It causes hemolysis by disrupting
the selective permeability of the
RBC membrane.
 Streptococcal Serology
▪ Infections and Sequelae:
1. Skin infections caused by S. pyogenes include:
a. Cellulitis b. Impetigo c. Erysipelas
2. Upper respiratory tract infections caused by S. pyogenes:
a. Sore Throat b. Pharyngeal Edema
3. Scarlet Fever
4. Rheumatic fever (RF)
5. Post-streptococcal Glomerulonephritis
Streptococcus pyogenes
 Streptococcal Serology
▪ Laboratory Diagnosis:

1. Anti-streptolysin O (ASO) Titer

2. Anti-DNAse B
3. Streptozyme Testing
 Streptococcal Serology
Laboratory Diagnosis:
1. Anti-streptolysin O (ASO) Titer
▪ The ASO titer begins to increase approximately 7 days after
infection and peaks after 4 to 6 weeks.
▪ Principle: (NEUTRALIZATION) SLO is added to serial
dilutions of patient serum, along with group O RBCs as
indicator cells.
▪ ASO titer is reported as the reciprocal of highest dilution
that shows no hemolysis & is expressed in Todd units.
Based on the neutralization of the hemolytic activity of streptolysin O
• Patient serum mixed with varying dilutions of standard amounts of
reduced streptolysin O antigen; mixture is added with standard
quantity of 5% suspension of rabbit or human red cells as indicator,
reincubated; presence of hemolysis is noted
• Tube containing the least amount of serum which completely inhibits
hemolysis represents the anti-streptolysin O titer if that serum
 Streptococcal Serology
Laboratory Diagnosis:
1. Anti-streptolysin O (ASO) Titer
▪ Normal Values:
Healthy adults have ASO titers of less than 166
Todd units, with the usual titer decreasing
after 50 years of age.
A 30% rise in titer above a previous level is of
greater significance than a single titer.
 Streptococcal Serology
Laboratory Diagnosis:
2. Anti-DNAse B (AD-B)
▪ Streptococci produce the enzyme Deoxyribonuclease B
(DNAse B).
▪ The anti-DN-B test is a neutralization test that can
demonstrate recent streptococcal infection.
▪ Anti-DN-B neutralizes the activity of DNAse B.
▪ Anti-DN-B levels are increased in the 15% to 20% of RF
patients who do not have elevated ASO titers.
• STREPTOZYME: SLIDE AGGLUTINATION SCREENING TEST FOR
DETECTION OF ANTIBODIES TO SEVERAL STREPTOCOCAL ANTIGENS.
• SHEEP RBC’S COATED W/ STREPTOLYSIN, STREPTOKINASE,
HYALURONIDASE, DNAse, so that ANTIBODIES to any streptoccal
antigen can be detected.
• Postive result : Hemagglutination
Anti-Streptolysin O (ASO) Latex Agglutination Test

• The Streptolysin O Antigen is fixed on the surface of the latex

particles. These latex particles are mixed with the patient’s
diluted serum on a slide and rotated. If the latex particles
agglutinate, the test result is positive. If there is no
agglutination, the test result is negative, if the qualitative
test result is positive, it can be quantitated to determine the
amount of ASO present.
Febrile Disease Serology
 Febrile Disease Serology
A. Febrile diseases are a group of microbial infections characterized by
fever and the production of antibodies known as febrile agglutinins.

B. These diseases include:

1. Brucellosis (Brucella abortus)
2. Paratyphoid Fever (Salmonella paratyphi)
3. Rocky Mountain Spotted Fever (Rickettsiae)
4. Tularemia (Francisella tularensis)
5. Q Fever (Rickettsiae)
A. SALMONELLA INFECTION: TYPHOID FEVER

• MOT: Ingestion of contaminated food or food products, or through

contaminated hands
• Infections include:
o Gastroenteritis (S.typhimurium, S.enteritidis)
o Bacteremia and Extraintestinal Infections (S.cholerasuis, S.dublin)
o Enteric Fever (Typhoid fever) (S.typhi, S. paratyphi)
• characterized by prolonged fever and multisystem involvement,
including lymph nodes, liver, and spleen
SALMONELLA ANTIGENS
Hauch or H antigen
• Thermolabile, Flagellar antigen
• Antigens are SPECIFIC for the given species
• Prepared by suspending the bacterial growth in saline containing 2%
formalin. Concentrated stock antigen is good for months, if
refrigerated
• Produce floccular type of agglutination
Ohne or O antigen
• Thermostable, Somatic antigen
• Directly associated with the bacterial body
• NON-SPECIE SPECIFIC and can divide genus into five groups (A,B,C,D,E)
• These are used to detect somatic agglutinins against Salmonella and
Proteus (OX2, Ox19, Oxk)
• Prepared by extracting bacterial cultures with either phenol or alcohol.
Concentrated stock antigen is good for months, if refrigerated
• Produce granular type of agglutination
Kapsel or K antigen
• Thermolabile, capsular antigen
• Occur as capsules or as envelop surrounding the bacterial body
• Varieties include: B, L, Vi antigens
• Vi antigen – occurs in highly virulent strains of Salmonella typhi,
S.paratyphi,and S.ballerup
• Also suggested as a means of identifying typhoid carriers who often
have negative “O” and “H” titers
 Febrile Disease Serology
C. Tests for febrile diseases include:

1. Widal’s Test, which can detect antibodies in typhoid fever,

tularemia and brucellosis.

2. Weil-Felix Test, which is an agglutination test based on the

cross-reactivity of rickettsial antibodies with antibodies to
the somatic “O” antigens of the OX-19 and OX-2 strains of
Proteus vulgaris and the OX-K strain of Proteus mirabilis.
 Febrile Disease Serology
Proteus Suspension
Infection
OX-2 OX-19 OX-K
Epidemic Typhus + + 0
Murine Typhus + + 0
Spotted Fever + + 0
Scrub Typus 0 0 +
Q Fever 0 0 0
Rickettsial Pox 0 0 0
Serological Tests for Other Bacterial
Infections
SEROLOGY OF VIRAL INFECTION

Hepatitis Virus
HIV
Epstein-Barr Virus (EBV)
Dengue Virus
Hepatitis Virus
 Hepatitis
“Inflammation of the Liver”
CAUSED BY: PRIMARY HEPATITIS VIRUSES
CHEMICALS DRUGS
AUTOIMMUNE DISEASES
 Hepatitis A
Infectious Hepatitis is caused by the HAV - PICORNAVIRIDAE
1.Transmission: Fecal Oral Route
2.Incubation Period: 28 Days
3.Disease Course: Acute and Self-limiting;
There is no carrier state.
4.Laboratory Diagnosis. Liver Function Test (ALT)Total Bilirubin
Antibodies to HAV can be detected by
Enzyme Immunoassay (EIA) and RIA
Methods.
 Hepatitis A
Infectious Hepatitis is caused by the HAV - PICORNAVIRIDAE
HAV ANTIBODIES
▪ Marker of Acute Hepatitis A
▪ Peak: During First Mons. of Illness
IgM Anti-HAV
▪ Declined: 6 to 12 Mons.
▪ Solid-Phase Antibody Capture ELISA
▪ Result of Natural Infection or
IgG Anti-HAV Immunization.
▪ Competitive Inhibition ELISA Test
 Hepatitis A
Infectious Hepatitis is caused by the HAV - PICORNAVIRIDAE

Prevention: Household and sexual contacts of infected

persons should receive immune globulin
injections within 2 weeks of exposure.
 Hepatitis B
Serum Hepatitis is caused by the HBV – HEPADNAVIRIDAE
formerly known as the Australia or
Hepatitis-associated Antigen
1.Transmission: Sexual Contact, Blood
2.Incubation Period: 60 to 90 Days
3.Disease Course: May be acute, chronic, or fulminant, or
the patient may be a chronic
asymptomatic carrier. Symptoms are
similar to those seen in HAV infections.
 Hepatitis B
Summarizes order of appearance of HBV Markers
 HEPATITIS B VIRUS ANTIGEN
▪ Australian Antigen.
▪ First Marker to appear.
Hepatitis B Surface Antigen
▪ Indicator of active infection
(HBsAg) ▪ Important marker in screening blood
donor.
▪ Present during active replication of
Hepatitis B Envelope Antigen
the virus.
(HBeAg) ▪ Indicates high degree of infectivity.
Hepatitis B Core Antigen ▪ Not detected in serum.
(HBcAg) ▪ Detected through Liver Biopsy.
 HEPATITIS B VIRUS ANTIBODIES
▪ Indicator of current or recent infection.
IgM Anti-HBc ▪ Detects the “Core Window Period”
IgG Anti-HBc ▪ Persists for the lifetime of the individual.
Anti-HBe ▪ Marker of Convalescence.
▪ Appears during the recovery period of Acute
Hepatitis B, weeks to months after HBsAg
Anti-HBs disappear.
▪ Provide Protective Immunity:
≥ 10 mIU/mL of Serum
HEPATITIS PROFILE RESULT & INTERPRETATION
ACUTE HEPATITIS A IgM anti-HAV (+)
RECOVERY FROM
Total anti-HAV (+)
HEPATITIS A
HBsAg (+)
Total anti-HBc (+)
ACUTE HEPATITIS B
IgM anti-HBc (+)
Anti-HBs (–)
 HEPATITIS PROFILE RESULT & INTERPRETATION
HBs Ag (–)
RECOVERY FROM HEPATITIS B Total anti-HBc (+)
Anti-HBs (+)
HBsAg (+)
Total anti-HBc (+)
CHRONIC HEPATITIS B/ CARRIER
IgM anti-HBc (–)
Anti-HBs (–)
HBsAg (–)
HEPATITIS B IMMUNIZATION Anti-HBc (–)
Anti-HBs (+)
HbsAg Anti Hbs Anti-Hbc STATUS ACTION
+ - +
- + +
- + -
+ - -
- - +
- - -
HEPATITIS SEROLOGICAL PROFILE
 Hepatitis B
Serum Hepatitis is caused by the HBV – HEPADNAVIRIDAE
Prevention: Avoidance of high-risk behavior (e.g., intravenous
drug abuse and sexual contact with infected
persons).
A vaccine against HBV has been available since
1982. In a health care setting, HBV vaccination
and the use of universal precautions can greatly
reduce the risk of occupationally acquired HBV.
TESTS AVAILABLE FOR HBV DETN
1st Generation Test
• 1. Ouchterlony
• Principle: Precipitation
2nd Generation Tests
• 1. Counter Immunoelectrophoresis (CIE)
• Principle: Precipitation with Current
• 2. Rheophoresis
• Principle: Precipitation by Evaporation
• 3. Complement Fixation
3rd Generation Tests (Most Sensitive)
1. Reverse Passive Latex Agglutination
• Principle: Agglutination
-antiHBsAg artificially/passively attached to latex particles
2. Reverse Passive Hemeagglutination
• Principle: Hemeagglutination
-antiHBsAg passively attached to red cells
3. ELISA
4. RIA
Other Hepatitis Virus
Human Immunodeficiency Virus
Human Immunodeficiency Virus
33.2 million people were living
with HIV infection, 2.5 million
people became newly infected
and 2.1 million people died of
AIDS
– WHO, 2007
 Human Immunodeficiency Virus
❑ Member of the Family: Retroviridae
Genus: Lenti virus

❑ Etiologic agent of Acquired Immunodeficiency

Syndrome (AIDS)

❑ Diameter of 100-120 nm with a spherical morphology

 Two Serotype
a. HIV-1
- Three subtypes
1. M
- Clades(A,C, D, H, G, K, F1, F2,J)
- Clade B (Homosexual)
- Clade A, C, E (Asia & Africa)
2. N (Non-M, Non-O)
3. O (Outlier)
b. HIV-2
- Subtypes (A-E)
 Viral Genome

Composed of 9 genes encoding 3 structural, 2 envelope, and 6

regulatory proteins
 ▪ Key Component for Viral Entry
Viral Replication
gp120 and gp41 – Attach to CD4

Primary Receptor:
- CD4

CORECEPTOR
- CXCR4
- CCR5

▪ Terms
a. T – tropic or X4 strains
b. M – tropic or R5 strains
c. Provirus
 MODES OF TRANSMISSION
1. Intimate sexual contact
2. Contact with BLOOD and other body fluid (SEMEN,
VAGINAL SECRETION, CSF, SYNOVIAL FLUID,
PERICARDIAL FLUID, PLEURAL FLUID, PERITONEAL
FLUID,AMNITIOC FLUID AND OTHERS)
3. Perinatally, from infected mother to infant
 SIGNS AND SYMPTOMS
 After exposure to HIV, some people have a flu-like illness
that lasts between a week to a month.
▪ Fever
▪ Headache
▪ Enlarged lymph nodes
 Several symptoms of occur due to a decreasing CD4 T cell
count including:
▪ Fatigue, weight loss
▪ Frequent fevers and sweats
▪ Persistent skin rashes or yeast infections
▪ Short-term memory loss
Symptoms of opportunistic infections:
▪ Coughing, shortness of breath
▪ Fever
▪ Lack of coordination, forgetfulness,
▪ Vision loss
▪ Persistent diarrhea
▪ Severe headaches
▪ Extreme fatigue
▪ Nausea, abdominal cramps, vomiting
▪ Conjunctivitis, ear infections, tonsillitis (children)
 COMPLICATION
▪ Kaposi’s sarcoma
▪ Cervical cancer
▪ Pneumocystis carinii : pneumonia
▪ Toxoplasma gondii, Cryptococcus neoformans
▪ CMV, HSV, Mycobacterium avium,
▪ Candida albicans, etc.
▪ Non-Hodgkin lymphoma
a. AIDS-related Burkitt lymphoma: chromosome-
translocation
b. AIDS-related Large cell lymphoma:
c. EBV infection
d. AIDS-related Primary effusion lymphoma: HHV-8 infection
 TREATMENT
❑ RETROVIRAL DRUGS

❑ PROPHYLACTIC THERAPY WITH

RETROVIRAL DRUG

❑ VACCINE
LABORATORY TESTING FOR HIV INFECTION
❑ HIV SCREENING TEST
1. ELISA
2. RAPID TEST (IMMMUNOCHOMATOGRAPHIC)
❑ HIV CONFIRMATORY TEST
1. WB
2. IFA
3. NAAT
❑ TEST STAGE AND MONITOR HIV
1. Viral Load
2. CD4 T-CELL COUNT
LABORATORY TESTING FOR HIV INFECTION
❑ HIV SCREENING TEST
1. ELISA
2. RAPID TEST (IMMMUNOCHOMATOGRAPHIC)
❑ HIV CONFIRMATORY TEST
1. WB
2. IFA
3. NAAT
❑ TEST STAGE AND MONITOR HIV
1. Viral Load
2. CD4 T-CELL COUNT
 Screening Test:
ELISA Test (Enzyme Linked
Immunosorbent Assay)

Confirmatory Test:
Western Blot or
Immunoflouroscent Assay
Appearance of HIV Markers
Appearance of HIV Markers
HIV Screening Test
False Positives and Negatives with HIV-
Antibody ELISA Testing
HIV Confirmatory/Supplemental Tests
Western Blot
Tests to Stage and Monitor HIV
Epstein-Barr Virus
 Epstein-Barr Virus (EBV)
▪ Causative agent of: Burkitt’s Lymphoma
Nasopharyngealcarcinoma
Infectious Mononucleosis (IM)

▪ The virus is ubiquitous; 80% to 90% of healthy adults have EBV

antibodies. EBV infects B lymphocytes.
 Epstein-Barr Virus (EBV)
Infectious Mononucleosis (IM)
▪ is an acute, self-limiting disease typically seen in young adults.
▪ The disease is characterized by fever, sore throat, cervical
lymphadenopathy, splenomegaly, and mild hepatitis.
▪ The WBC count is elevated, and reactive lymphocytes are
seen in the peripheral blood.
▪ There is a relative and absolute lymphocytosis.
▪ The average incubation period is approximately 2 to 8 weeks.
 Epstein-Barr Virus (EBV)
Antigens and Antibodies:

1.VIRAL CAPSID ANTIGEN (VCA) is found in the cytoplasm of EBV-

infected lymphocytes. IgM antibodies against VCA are detectable
early in the infections, but disappear within 2 to 4 months. IgG
antibodies against VCA develop within 1 week after infection and
can persist for life.
 Epstein-Barr Virus (EBV)
Antigens and Antibodies:

2. Early antigen-diffuse (EA-D) and early antigen-restricted (EA-R)

antigens are found in the cytoplasm of infected B lymphocytes.
EA-D is also found in the nucleus. IgG antibodies to EA-D can be
indicators of active disease. IgG antibodies to EA-R are
sometimes seen in young children who have active IM infection,
but not in infected young adults.
 Epstein-Barr Virus (EBV)
Antigens and Antibodies:

3. Epstein-Barr nuclear antigen (EBNA) is found in the nuclei of all

infected cells. IgG antibodies to EBNA develop slowly but can
remain detectable throughout life.
 Epstein-Barr Virus (EBV)
Antigens and Antibodies:

4. Heterophile antibodies are stimulated by one antigen and will

react with unrelated antigens from different mammalian
species. The heterophile antibodies of IM are IgM antibodies
and are seen in 50% to 70% of patients with IM. They persist for
4 to 8 weeks after infection.
• IgM anti VCA- most useful marker for acute IM
• IgG anti –VCA present at the onset of IM, persisit for life (which
indicates past infection)
• Anti EBA- appears during convalescence
 Epstein-Barr Virus (EBV)
Serological Testing:

The Paul-Bunnell Test

▪ Can detect only the presence or absence of
heterophile antibodies.
▪ It cannot determine the specificity of the antibodies.
 Epstein-Barr Virus (EBV)
Serological Testing:

The DAVIDSOHN DIFFERENTIAL TEST can distinguish heterophile

sheep cell agglutinins in human serum caused by IM, serum
sickness, and Forssman antigen.

MONOSPOT TEST is based on the principle that horse RBCs are

agglutinated by the heterophile antibodies of IM.
 Davidson Differential Test
Adsorption Pattern
Type of Absorption by
Absorption by Beef
Heterophile Guinea Pig Kidney
RBCs
Antibody Tissue
Antibodies in IM - +
Forssman + -
Serum Sickness + +
 Davidson Differential Test
Agglutination with Sheep RBC’s after Adsorption
Type of Absorption by
Absorption by Beef
Heterophile Guinea Pig Kidney
RBCs
Antibody Tissue
Antibodies in IM + -
Forssman - +
Serum Sickness - -
Serologic Test for Diagnosis of Recent Infection
Serologic Test for Diagnosis of Recent Infection
Clinically
Organism Test
Significant Result
ASO ≥ 1:240
S. pyogenes Anti-DNAse B ≥ 1:240
Anti-Hyaluronidase ≥ 1:512
S. typhi Widal Test ≥ 1:160
L. pneumophila Indirect IFA ≥ 1:256
RPR (+)
T. pallidum VDRL (+)
FTA-ABS (+)
Serologic Test for Diagnosis of Recent Infection
Clinically
Organism Test Significant
Result
B. burgdorferi Indirect IFA ≥ 1:64
EIA (+)
Western Blot IgG ≥ 4 of 9 bands
Western Blot IgM ≥ 2 of 9 bands
H.pylori EIA (+)
Serologic Test for Diagnosis of Recent Infection
Clinically
Organism Test Significant
Result
Cold Agglutinins ≥ 1:128
M. Pneumoniae Complement Fixation ≥ 1:32
EIA (+)
R. rickettsi Indirect IFA ≥ 1:64
E. chaffeensis Indirect IFA ≥ 1:64
B. henselae Indirect IFA ≥ 1:128
Serologic Test for Diagnosis of Recent Infection
Clinically
Organism Test Significant
Result
Indirect Hemagglutination ≥ 1:256
E. histolytica
EIA (+)
Indirect IFA ≥ 1:64
T. gondii
EIA (+)
Indirect Hemagglutination ≥ 1:128
Cysticercosis
EIA (+)
Bentonite Flocculation ≥ 1:5
T. spira
EIA (+)
Toxocara sp. EIA ≥ 1:32
Serologic Test for Diagnosis of Recent Infection
Clinically
Organism Test Significant
Result
Immunodiffusion (+)
Aspergillus sp.
Complement Fixation ≥ 1:32
Immunodiffusion (+)
B. dermatitidis
EIA ≥ 1:32
Immunodiffusion (+)
Candida sp.
Latex Particle Agglutination ≥ 1:80
Immunodiffusion (+)
H. capsulatum
Complement Fixation ≥ 1:32
Test/ procedure Parasite infection detected
String or Entero test G. lamblia, S. stercoralis (Beale’s string test)

Culture in Diamond or Feinbergh and Whittington’s T. vaginalis

medium

Culture in Novy-MacNeal-Nicole medium Leishmania and Trypanosama

Montenegro intradermal test L. tropica, L. braziliensis
Xenodiagnosis T. cruzi, T. spiralis
Sabin-Feldman dye test T. gondii
Autoflourescence C. cayatenensis
Parasight F test P. falciparum
Casoni’s intradermal test E. granulosis
Circumoval precipitin S. japonicum
Bachman intradermal T. spiralis
Bentonite flocculation test