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28 Cards in this Set
- Front
- Back
- 3rd side (hint)
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Difference between Post/Pro zone |
Post zone - excess of antigen Pro zone - excess of antibody |
PoG BRo |
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Refrigeration and Centrifugation time for Serial Dilution |
30 mins Refrigeration 20 secs Centrifugation |
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Reagents and other Solutions required for Serial Dilution |
0.25mL saline (per tube) 0.25mL antibody A / Anti-A Antisera 0.25mL of 3% RBC Suspension (per tube) |
NSS Aa RCS |
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Define Titer |
Last tube/dilution with agglutination signifying test's endpoint |
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Last step before adding the RBC suspension to each tube |
Discard 0.25mL from the last tube |
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Dilution Formula: |
Dilution = Amount of Solute / Total Sol'n Volume |
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Total Volume Formula: |
Total Volume = Amount of Solute + Diluent |
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Dilution Ratio Formula: |
Dilution Ratio = Amount of Solute / Total Volume |
DR = Sol't / T Vol. |
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How the get the ratio of dilution per tube? |
Multiply the dilution factor to the number of tube as coefficient factor.
Ie.: If first tube is 1/2, then 6th tube is?
1 - 1/2 2 - 1/4 3 - 1/8 4 - 1/16 5 - 1/32 6 - 1/64
n = (1/2)^6
n = 64 DILUTION RATIO = 1/64
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Prep of RCS requires what blood sample? |
Anti-coagulated blood in a Lav/EDTA Evacuated Tube |
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Purpose of RCS in Serial Dilution |
Indicator of titer via Agglutination |
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Process of RCS Prep: |
1. Centrifuge EDTA blood for 10mins. Discard the plasma. 2. Move remaining RBC to diff. tube; place at least 3/4 NSS into the tube . 3. Apply parafilm and Centrifuge for 1min. Decant the supertanant.. 4. After 3rd wash, pour 5mL NSS and 40uL if RCS to create ~5% suspension * 4mL NSS + 100uL RCS can also be use |
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Correctly prepped RCS is characterized as: |
Tomato Red Appearance |
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Process that removes unbound antibodies in RCS |
RBC Washing |
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Buffer/Dilutor Solution for semi-quantitative serial dilution: |
Glycine Saline Buffer |
Gly Salt Bfr |
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Area in the tube with visible precipitation |
Zone of Equivalence |
ZoE |
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Dilution Technique for large quantities of samples |
Compound Dilution |
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Principle of the ABO Blood Typing Test |
Hemagglutination Reaction (RBC agglutination) |
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Materials of Blood Typing |
Reagents: Antibody A antisera Antibody B antisera Antibody D antisera (Rhesus Type)
Anti-coagulated Whole Blood Sample (EDTA/Lav Tap)
Slides, Pipettes, Applicator Stick |
ABDS
SPA
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Source of Error in reading Blood Type test: |
Prolonged drying/standing of Blood in slides |
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Agglutination Reaction in each Blood type (no Rh) |
(+) Agglutination occured between Antibody antisera and RBC Antigen
Type A (+) Anti-A
Type B (+) Anti-B
Type AB (+) Anti-A (+) Anti-B
Type O (-) Anti-A (-) Anti-B |
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Blood typing technique that pairs and Unknown blood Type with a known antibody for an antibody-antigen reaction to occur |
Forward/Direct Blood Typing |
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Agglutination processes |
1. Sensitization - rapid and reversible combination of AntiG-AntiB through single antigenic determinants
2. Lattice Formation - stable formation of AntiG-AntiB complexes in cross-links of multiple antigenic determinants, forming visible aggregates |
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Factors that affect Agglutination |
Epitopes : amount or obscured = less Antibody interaction
Charges of cellular components : Bacterial cells and RBCs may repel each other (both - charge) |
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IgM vs IgG in agglutination reaction |
IgM: - large, pentamer with more binding sites (epitopes) - best reacts at 4C to 27C
IgG: - require physicochemical conditions: soln' ionic strength, pH level, temp. - best at room temp. (30C to 37C) |
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Titer techniques with Semiquantitative results can be done in: |
Test Tubes or Microtiter Plates |
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Principle of Serial Dilution through Hemagglutination and its results |
Cell Sedimentation Pattern
(-) dark red, smooth button (+) cells in a jagged pattern with irregular edge |
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Enhancement Media for IgG Blood Typing |
22% Bovine Serum Albumin (30mins) Low Ionic Strength Solution (10-15mins) Polyethylene Glycol (not for patients with elevated proteins - ie. Multiple Myeloma) |
--% BovSAlb (__mins)
LISSol'n (__ to __ mins)
PoEthGly (Mul. Mye) |
