lecture 8

Question 1

detect antibodies through

Answer 1

infectious agents
vaccine for antigens (measure primary or memory responses)
autoantibodies: anti-nuclear antibodies
maternal antibodies

Question 2

reagent antibodies

Answer 2

allow us to detect antigens of pathogens in tissues

Question 3

cell surface antigens detected by antibodies tell us

Answer 3

blood types
lymphocyte subsets: lymphoma, leukemia, immunodeficiency
tumors and tumor markers in tissues and blood

Question 4

the specificity of antibody mediated immune responses can be exploited for

Answer 4

diagnostic purposes

Question 5

serology confirms

Answer 5

results of other methods (virus isolation, PCR etc)

surveillance of infections in herds (detect antibody or monitor antibody levels)

Question 6

serology

Answer 6

test of choice for many agents for which no test is available

Question 7

test results from any serological test means

Answer 7

there has been an exposure to pathogens and/or some protein (components of pathogens as vaccine - antibodies present bc there was vaccine)
in young animal it also means they could receive antibodies from mother

Question 8

serum test sample

Answer 8

top fraction after blood sample clots
contains serum proteins such as antibodies and many other proteins

antibodies will tell what antigens present in body

Question 9

body cavity fluids test sample

Answer 9

peritoneal fluid: feline infectious peritonitis (FIP) virus

cerebral spinal fluid: west nile virus (WNV - TLR3 issue)

Question 10

whole blood in anticoagulant

Answer 10

contains RBC, WBC and platelets

ex: coombs’ test, flow cytometric immunophenotyping

Question 11

tissue sections

Answer 11

necropsy or surgical biopsy specimens

ex: immunohistochemistry or immunofluorescence for bovine viral diarrhea virus (BVDV) FIP

Question 12

polyclonal antibodies

Answer 12

type of serologic reagent used in immunodiagnositics
multiple B cell clones make antibodies to different epitopes
antibodies generated against a certain pathogen or molecule
antiglobulin or antiserum consists of polyclonal antibodies made against immunoglobulins of different species

Question 13

isotypes and half lifes are important in

Answer 13

antibody test result interpretation

Question 14

monoclonal antibodies

Answer 14

type of serologic reagents used in immunodiagnostics
single B cell clone makes antibody to one epitope
made w hybridoma technology
hybridoma

Question 15

hybridoma

Answer 15

hybrid cell line generated by fusion of a B cell (ANTIBODY PRODUCER) w an immortal neoplastic B cell (myeloma) - bad

Question 16

monoclonal antibody production

Answer 16

host animal immunized w antigen of interest -> spleen lymphocytes harvested and fused w immortalized myeloma cells -> transfer to media w selection agent for immortalized hybridoma cells -> individual hybridoma cells are screened for antibody production -> cells that make antibody of desired specificity are expanded from a single antibody-producing clone

all descendent cells make the identical antibody

Question 17

major types of immunodiagnostic tests

Answer 17

immunofluorescene assay (IFA)
enzyme assay
precipitation assay
agglutination assay

Question 18

immunofluorescence assays (IFA)

Answer 18

antigen detection
antibody detection
serum or virus neutralization assay

can be direct or indirect

Question 19

enzyme assays

Answer 19

enzyme linked immunoabsorbant assay (ELISA) - indirect
immunohistochemistry
western blot

Question 20

need to wait how many days before can detect antibody in serum?

Answer 20

14 days

Question 21

direct immunofluorescence

Answer 21

uses a primary antibody to detect the target protein

Question 22

indirect immunofluorescence

Answer 22

uses antibodies in patients serum (PRIMARY ANITBODY) to detect an antigen followed by a secondary antibody to detect the primary antibody

Question 23

ELISA

Answer 23

enzyme linked immunosorbent assay
enzyme assay - indirect

absorbance is proportional to the amount of antibody attached to the antigen and proportional to the amount of antigen attached to well

Question 24

SN

Answer 24

serum neutralization

Question 25

HI

Answer 25

hemagglutination inhibition assay

Question 26

IFA

Answer 26

indirect fluorescent asay

Question 27

CF

Answer 27

complement fixation

Question 28

WB

Answer 28

western blot | enzyme assay

Question 29

AGID

Answer 29

agar gel immunodiffusion assay

Question 30

IFA indirect test results

Answer 30

positive (antibody is present in serum) is fluorescent fluorophore binds to antibody (Ig) in patient serum and will cause stain antiPCV antibody test can use other animal to produce antibody to virus that would bind to PRRS virus in mouse? detects antibodies

Question 31

antiPCV antibody test (ANA)

Answer 31

used in the diagnosis of immune response to PCV2 or any other viruses IFA

Question 32

serum neutralization test

Answer 32

tests function of the patient antibody, whether or not its antibodies block the virus infection or not antibody present in serum + virus (incubation) -> target cell not infected - no fluorescence (FAVN) no antibody present in serum + virus -> specific monoclonal antibody binds to flurophore and shows flourescence

Question 33

FAVN

Answer 33

fluorescent antibody virus neutralization test | serum neutralization test

Question 34

ELISA indirect test

Answer 34

well is precoated w antigen -> test serum is added w chosen antibodies to test for -> labeled antiglobulin is added -> enzyme substrate added and will wash away any nonbound antibody -> color changes when bound

Question 35

ELISA competitive test

Answer 35

no antibodies = color = negative antibodies present = no color = positive labeled and unlabeled (patient serum) antibodies compete for binding to antigen -> addition of enzyme substrate leads to a color change that is inversely proportional to amount of patient serum (monoclonal antibodies!) antibody bound more accurate

Question 36

ELISA sandwich test

Answer 36

well is precoated w capture antibody -> test antigen is added -> detection antibody is added -> labeled antiglobulin is added -> color changes after enzyme substrate is added antigen is bound to plate by means of an antibody (captured antibody!) presence of bound antigen is detected by sequential addition of a secondary antibody (detection antibody) and an enzyme labeled antiglobulin addition of enzyme substrate leads to color change proportional to amount of bound antigen color = positive!!