MTAP 1 10/15/2022 Flashcards
gel test technology was developed by
Dr. Yves Lapierre in 1985
gel test technology: gel type
dextran polyacrylamide gel
gel test technology
microtube upper portion is
reaction of chamber
gel test technology
microtube lower portion is
column
gel test technology principle
size exclusion chromatography
type of gel tests
- specific
- plain / neutral
- low ionic antiglobulin
type of gel test specific for target antibody/antigen
specific
Type of gel test
- The gel is not yet dispensed with reagent
- Add the reagent along with the sample in the microtube
Plain / Neutral
gel test that do Reverse/Backward/Indirect/Serum Typing
plain / neutral
test gel specific for sensitization detection
low ionic antiglobulin
low ionic antiglobulin reagent added into the matrix
AHG reagent
Agglutination reaction in gel medium can be reviewed for _ DAYS (clump is stable)
3
gel test technology sample volume
<1ml
gel test:
The gel matrix itself traps the unbound _ that may interfere with the reaction
antibodies
grading of RBC most cells at the center portion in gel test
2+
grading of RBC near the bottom portion in gel test
weak (+/-)
antihuman globulin test is also known as
Coombs Test
end product of lattice formation
agglutination
best agglutinating antibody
IgM
Incomplete IgG antibodies are only capable of _ the red cell antigens.
sensitizing
principle of antihuman globulin test
detection of cell-bound immunoglobulin / incomplete antibodies (IgG)
cold reacting antibody is best reactive at what temperature?
1-6deg C
saline-reactive immunoglobulin
IgM
albumin / ahg reactive immunoglobulin
IgG
IgM ab example
IgG ab example
ABO
Rh
IgG Most to least potent:
3,1,2
type of AHG reagents
polyspecific AHG reagent
monospecific AHG reagent
polyspecific ahg reagent is consist of
- pool of rabbit anti-human IgG
- mouse monoclonal anti-C3d and anti-C3b
Polyspecific AHG Reagents are also refereed to
Broad Spectrum Coombs Reagent
Add _ to distinguish AHG from other different reagent
green dye
Method used for Monospecific reagent
Hybridoma Technique/Kohler and Milstein Technique
cells combined in forming hybridoma cells
mouse plasma cells and myeloma cells
stages of ab-ag interaction
- sensitization
- agglutination
DAT detects _ sensitization
in vivo
Immediate HTR is severe and fatal and usually
caused by
ABO incompatibility
DAT collection tube
EDTA and citrate
chelation of calcium holds _ subunits (_) together
C1; q,r,s
largest complement protein
c1
wharton’s jelly can cause
rouleaux formation
presence of unbound antibodies can _ the AHG reagent resulting to
neutralize; false negative
Delay in the addition of reagent = antibodies may
undergo
ELUTION PROCESS
conventional method of DAT
- washing
- ahg addition
- centrifugation
- dislodge
- observe
minimum ration of serum to cells in AHG Test
40:1
in AHG Test, drops of serum and cells
2 drops serum and 1 drop of 5% v/v cell suspension
too high temperature in AHG Test can cause
protein denaturation
too low temperature in AHG Test can cause
IgG inactivation
in AHG Test incubation time in saline suspension
30-120 minutes
in AHG Test incubation time in LISS suspension
10-15 minutes
in AHG Test saline for washing; should be fresh and buffered to a pH of _
7.2-7.4
in AHG test centrifugation time
1000 rcf for 15-20 seconds
