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immuno > techniques > Flashcards

techniques Flashcards

(53 cards)

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1
Q

agglutination

A

reactions between insoluble and soluble component

-takes place on the surface of something

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2
Q

2 main steps of agglutination

A

sensitization
lattice formation

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3
Q

sensitization stage

A

antigen and antibody combined through antigenic determinant sites

-rxn is rapid and reversible

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4
Q

lattice formation

A

crosslinking that form visible agglutination you see

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5
Q

too much antigen

A

post zone

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6
Q

too much antibody

A

pro zone

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7
Q

how to manipulate surface charge

A

add LIS - low ionic strength saline
albumin
enzymes

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8
Q

drawback when testing for IgM

A

rheumatoid factor

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9
Q

direct agglutination

A

antigens found on a particle

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10
Q

hemagglutination

A

particle involved is a RBC

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11
Q

passive/ indirect agglutination

A

particles are coated with antigen that is normally not found on them

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12
Q

reverse passive

A

antibody is coated on a particle

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13
Q

coagglutination

A

bacteria is inert particle to which a specific antibody is bound

-looking for antigen in patient serum

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14
Q

most common coagglutination

A

staph aureus – stick to protein A and look for presence of Antigen

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15
Q

one of first method used for hcg in urine

A

Agglutination inhabitation

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16
Q

how does Agglutination inhabitation work

A

negative reaction (no agglutination) is actually positive

-add patient serum (with antigen) add this to an Ab

if Ag is present on = Ag-ab response
-readd Ab coated with antigen– no attachment so negative rxn

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17
Q

what test use flocculation

A

RPR and VDRL (non-treponemal )

-this is a form of precipitation

antigen is lipid in nature– loose aggregates

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18
Q

precipitation

A

coming together of 2 soluble components to form insoluble complex

-less sensitive than agg

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19
Q

what type of rxn is precipitation

A

Secondary type reaction – because dependent on Ag-Ab coming together and then precipitating

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20
Q

affinity

A

initial attraction

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21
Q

avidity

A

strength of ag-ab

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22
Q

turbidity measurements

Study These Flashcards
A

often measured how light is scattered in a spec

measures: amount of cloudiness (Ag-ab response)

no Ag-ab response= a lot of light coming through

23
Q

nephelometry

Study These Flashcards
A

Light scattered at an angle proportional to amount of Ag-Ab in a sample

-more complex= more scatter

detector at 90, 45 and straight line

24
Q

what is beer’s law

Study These Flashcards
A

Relationship between absorption of light with a solution and the concentration of the solution

-used in photometric methods

25
in beer's law, absorption is proportional to
concentration if certain parameters are met A1C1= A2C2 reverse is A=-log%T
26
slope in beer's law
1 -works good when absorbance is less than 2
27
radial immunodiffusion
single diffusion tech. -only 1 component moves
28
what does Radial Immunodiffusion (RID) measure
immunoglobulins or complement components
29
procedure for Radial Immunodiffusion (RID)
- Antibody in a support gel -RBCs added to gel -- give macroscopic appearance of how antigen moved - Add patient serum (Ag) - Look at diameter of ring formed - more antigen larger the zone size
30
End Point- Mancini method
takes 48-72 hours
31
kinetic or fahey method
-faster but still long need to find line of best fit measurements on semi log paper
32
what needs to run for kinetic method
Need 3 controls run and then run patient
33
graph specific for kinetic method
x=diameter y= concentration measure diameter and plot graph
34
ex of label immunoassays
ELISA Fluorescent
35
why do we like label immunoassays
* Quick, can be automated * Reproducible * Depend on affinity and avidity -Most good sensitivity and specify
36
enzyme immunoassays either
competitive or noncompetitive assay
37
more common enzyme immunoassay
non competitve
38
competitive assay
* Labeled Ag competing with unlabeled antigen for specific number of binding sites * Amount of bound labeled component is inversely proportional for the amount of antigen you are looking for
39
in competitive If measured labeled component is low --
a lot of antigen from the patient
40
typical ELISA procedure is
noncompetitive -high sensitivity
41
most common anti-ab
alkaline phosphatase or horseradish peroxidase -stable and won't interfere with testing
42
most common chromagen
P-nitrophenyl phosphate
43
capture assay what kind
Noncompetitive labeled assay
44
what is bound to solid particle in capture assay
antibody -you are detecting the Ag
45
capture assay is best used for
antigens that are multivalent -IgM
46
what can interfere with capture assay
rheumatoid factor
47
why is capture assay different from ELISA
Ab is bound to solid particle and Ag is captured
48
lateral flow immunoassay also known as
immuno chromogenic assay
49
what is lateral flow called what it is
components laterally flow down cassette (precipitation reaction)
50
can you dilute antigen or antibody in lateral flow
no
51
drawback of fluorescent tech
-need fluorescent scope -are you reading the correct component or is it background noise
52
2 kinds of fluorescent tech
Direct : looking for Ag Indirect : looking for Ab
53
what is the tag in fluorescent
FITC- fluorescent isothiocyanate

immuno (12 decks)

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