Serology Day 2 - Serological techniques, TORCH, syphilis

Question 1

Immunofixation Electrophoresis

Answer 1

Serum run on different rows of gel, anti-antibodies used to see each type of immunoglobulins in each row

Question 2

Titer results

Answer 2

Dilution factor of last dilution that shows positive result

Question 3

Pre-zone

Answer 3

More antibodies than antigen

Question 4

Post-zone

Answer 4

More antigen than antibody

Question 5

Zone of equivalence

Answer 5

Antigen and antibody concentrations close enough to form immune complexes

Question 6

Precipitation assays

Answer 6

soluble antigen reacts with soluble antibody (turbidity/nephelometry)

Question 7

Ouchterlony formation patterns

Answer 7

Identity: solid bent line between antibody, unkown antigen, and standard antigen
Non-identity: crossed lines, little similarity between standard and unkown antigen
Partial identity: lines close but obviously distinct, standard and unknown antigen have similar epitopes

Question 8

Radial immunodiffusion

Answer 8

Agar has antibody in it, patient sample added to well, allowed to difuse
Ring is formed, diameter of ring is proportional to concentration

Question 9

Immunoelectrophoresis

Answer 9

Antigens run on gel first, trough cut and antibody added which diffuses

Question 10

Hypogammaglobulinemia

Answer 10

Low amounts of Ig’s

Question 11

A-gammaglobulinemia

Answer 11

Absence of Ig’s

Question 12

Hypergammopathy

Answer 12

Too many Ig’s

Question 13

Direct agglutination

Answer 13

Antigen directly combines with antibody

Question 14

Indirect agglutination

Answer 14

Carrier molecule coated with antigen, reacts with antibody in more visible way

Question 15

Hemmaglutination

Answer 15

ABO blood group assay

Question 16

Flocculation test

Answer 16

Precipitate of fine particles, use indirect method

Question 17

Complement fixation test

Answer 17

No hemolysis = complement reacted with Ag/Ab complex (Ab is present in patient)
Hemolysis = complement not fixed onto Ag/Ab complex, no Ab in patient

Question 18

Western Blot

Answer 18

Proteins, using electrophoresis the use anti-protein antibody

Question 19

Southern Blot

Answer 19

DNA, using electrophoresis and radioisotopes

Question 20

Heterophile antibodies

Answer 20

IgM’s produced in infection that are capable of binding to unrelated antigens from other species

Question 21

Forssman antibody

Answer 21

Antibody developed during mono infection also reacts against guinea pig kidney and sheep RBC

Question 22

Heterophile antibody tests

Answer 22

Paul Bunnell, Dadisohn Differential

Question 23

TORCH

Answer 23

Toxoplasmosis
Other
Rubella
Cytomegalovirus
Herpes

Question 24

Toxoplasma gondii

Answer 24

Asymptomatic to mono-like symptoms (birth defects cause hydrocephalus)
Domestic cat is host, difficult to culture, use EIA to test

Question 25

Cytomegalovirus

Answer 25

Part of herpes family (stays latent mostly, mild mono-like symptoms when reactivates), almost everyone has it (passed by fluids)

Question 26

Rubella

Answer 26

"3 day measles", serious birth defects like CHF, considered immune at 1:8 titer, 4 fold increase shows recent infection

Question 27

Herpes

Answer 27

Genital virus, can pass to baby during birth if mom is active, can kill infant

Question 28

Syphilis bacterium

Answer 28

Treponema pallidum (a spirochete)

Question 29

Syphillis (background info)

Answer 29

Can be destroyed by heat, cold, and drying Need direct contact with open lesion to pass on, can pass to infants during birth Treat with penicillin

Question 30

Syphillis clinical presentation

Answer 30

Primary: chancre (gential lesion) Secondary: malaise, asymptomatic Tertiary: inflammation, neurosyphillis

Question 31

Syphillis screening

Answer 31

Start with non-treponemal antibodies (reagin), then look for treponemal antibodies

Question 32

Nontreponemal tests

Answer 32

Venereal Disease Research Laboratory (VDRL) and Rapid plasma reagin (RPR)

Question 33

VDRL test

Answer 33

Looking for flocculation on slide, can do quantitative to monitor treatment, CSF can only be tested via VDRL

Question 34

RPR test

Answer 34

Looking for flocculation, sensitive, faster than VDRL

Question 35

Treponemal tests

Answer 35

Fluorescent treponemal absorption (FTA-ABS) and T pallidum particle agglutination (TP-PA)

Question 36

FTA-ABS test

Answer 36

Incubate with sorbent (remove cross-reacting antibodies), incubated on slide with killed T pallidum, use fluorescent anti-human Ig antibody to view

Question 37

TP-PA test

Answer 37

Patient serum incubated wit gel particled covered in T pallidum antigen, smooth mat on surface of well is positive rxn, button is negative rxn

Question 38

Typical antibody patterns in syphillis

Answer 38

Treponemal antibodies stay high throughout all stages, even after treatment Nontreponemal antibodies decrease as disease become latent (treated drops lower than untreated)