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45 Cards in this Set
- Front
- Back
- 3rd side (hint)
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Precipitation vs. Agglutination
Interaction |
Interaction between Ab and Ag vs. Interaction between Ab and particulate Ag
Location and observation |
large, insoluble complex in a solution vs. Large aggregates or clumps |
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Agglutination
Ags must be? |
Particulate or on a cell surface |
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Ouchterlony
Procedure |
Ag and Ab are placed in agarose well |
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Ouchterlongy
Method |
Double diffusion |
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Ouchterlongy
What is being compared? |
Reactivity between? |
2 Ags and 1 Abs 1 Ags and 2 Abs |
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Ouchterlongy
It is used to identify? |
unknown
Ag or Ab
in? |
Pt. sample |
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Ouchterlongy
3 patterns of precipitation |
Identity Nonidentity partial Identitiy |
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ocuhterlongy
__________ Nonidentity Partial identitiy |
Identity
Definition |
Ags are recognized as identical by Ab |
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Ouchterlongy
Identity Ags are recognized as identical by Ab
Observation |
2 rxn fuse together to form an arc |
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Ouchterlongy
Identity Example: Ab 1 + Ag 1 (known reagent) + Pt serum (unknown)
Meaning |
Pt serum contains Ag1
Observation |
Arch |
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Ouchterlongy
Non-identity
Def. |
Ags are recognized as nonidentical |
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Ouchterlongy Non-identity
Observation |
X Pt line cross over each other
Result from? |
2 separate rxn |
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Ouchterlongy Non-identity
Ag1 (known reagent) Pt serum Ab1 and Ab2
Conclusion |
Pt serum contains Ag 2 and no Ag1 |
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Ouchterlongy
Partial identity
Definition |
Ags are partially identical and share some of the epitopes
Related concept |
Cross-reactivity |
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Ouchterlongy
Cross reactivity |
Ab reacts with an Ag that is structurally similar to the original Ag
Notes |
Ab reacts an Ag that is structurally similar to the original ag that induced production of aB |
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Ouchterlongy
Partial identity
Observation |
Fusion of 2 lines with a spur
What does this spur point to? |
Least reacting or simpler ag |
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Ouchterlongy
Partial identity
Ag1: A and B Pt serum (Ag2): A and C Ab A and B
Conclusion |
Spur to the Pt serum
It has A but not B
See slide |
11 |
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Quantitative ouchterlony
How titer is determined? |
Last well with precipitation
What about the well with no precipitation? |
N |
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Latex Agglutination
How many types there are? |
2
List |
passive Reverse passive agglutintonation |
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Latex agglutination
______________ Reverse passive agglutination |
Passive agglutination
What is being detected? |
Ab in Pt. sample |
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Latex agglutination
Passive agglutination
Detects Ab in Pt. sample
procedure |
1. Pt. serum + Latex with Ag 2. Mix 3. incubate 4. Observation, what is being observed? |
Agglutination: clumping of latex beads |
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Latex agglutination
Passive agglutination
How the Ag is attached to the latex bead? |
Passively |
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Latex agglutination
Passive agglutination
Interpretation of results |
+ Ab: - Ab: |
+ Ab: agglutination - Ab: no agglutination |
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Latex agglutination
passive agglutination ______________ |
Reverse passive agglutination
What is being detected? |
Ag in pt. sample |
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Latex agglutination
Reverse passive agglutination
Ag is being detected
Procedure |
1. Pt serum + latex with ab 2. Incubate 3. Mix 4. Observation, what is being observed? |
Agglutination |
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Latex agglutination
Reverse passive agglutination
Interpretation of results |
+ Ag: - Ag: |
+ Ag: agglutination - Ag: No agglutination |
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Passive vs. reverse passive agglutination
What is being detected? |
Ab vs. Ag |
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Passive vs. reverse passive agglutination
What is passively attached to the latex beads? |
Ag vs. Ab |
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Hemagglutination
Def. |
Agglutination of RBCs
How many types there are? |
2 |
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2 types of hemagglutination |
Active hemagglutination Passive hemagglutination
Aka |
Direct Indirect hemagglutination |
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2 types of hemagglutination
__________ passive hemagglutination |
active hemagglutination
aka What is being detected? |
Presence of an Ab directed against a RBC ag (found naturally) |
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Active hemagglutination
Procedure |
1. Pt serum (________ and _____)+ RBC (____) 2. Incubate 3. Look for hemagglutination |
Pt. serum (serial dilution and Anti-RBC?)
Ag |
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Active hemagglutination
Interpretation of results |
+ Ab: - Ab: |
+ Ab: agglutination - AB: N |
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Active hemagglutination
Titer? |
Last tube that has agglutination |
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Application of active hemagglutination
Examples |
Infectious mononucleosis - Heterophile - Cold agglutinins
Review: what is being detected in active? |
presence of an Ab directed against a RBC Ag (Found naturally) |
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Passive (indirect) hemagglutination
Characteristic |
RBC serves as inert carrier of Ag |
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Passive (indirect) hemagglutination
RBC serves as inert carrier of Ag
Do Ags spontaneously adhere to RBC? |
Y, example?
and
N, Solution? |
Y: polysaccharides
N: chemically attached (tannic acid) |
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Passive hemagglutination
Procedures |
1. Pt serum (___ and ____) + RBC-Ag 2. Incubate 3. Look for agglutination |
Ab to Ag? serial dilutions |
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Passive hemagglutination
Interpretation of results |
+ Ab: - Ab: |
+Ab: agglutination -Ab: N |
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Passive hemagglutination
Titer |
Last tube with agglutination |
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Types of error in Agglutination rxns |
FP FN |
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Types of error in agglutination rxns
___ FN |
FP
Cause |
Drying out of reagents |
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FP in agglutination rxns
Drying out of agglutinations
Cause |
2
List |
Improper pipetting Delayed reading |
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Agglutinations rxns
FN
Cause |
Vigorous shaking of __________ and _______ |
test tubes microtiter plate |
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Agglutination rxns
FN
Cause: Vigorous shaking of test tubes or micropipette plate
Effects |
Disruption of complex formation |
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