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Rheumatoid Factor (Rf) Latex Slide Test

Rheumatoid Factor (Rf) Latex Slide Test

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Published by kiedd_04
The RF reagent is a suspension of polystyrene latex particles sensitized with specially prepared human IgG. The reagent is based on an immunological reaction between human IgG bound to biologically inert latex particles and rheumatoid factors in the test specimen.
The RF reagent is a suspension of polystyrene latex particles sensitized with specially prepared human IgG. The reagent is based on an immunological reaction between human IgG bound to biologically inert latex particles and rheumatoid factors in the test specimen.

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Published by: kiedd_04 on Oct 24, 2008
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04/30/2013

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[PRACTICAL 7]MTEB 2404
INDIRECT AGGLUTINATION ASSAY 
PRINCIPLE
Indirect agglutination test involves the use of inert agglutinableparticles (e.g. red cells or latex particles) as passive carriers for smallsoluble antigens or antibodies. In this case, the soluble antigens /antibodies are adsorbed to, or covalently linked to, the surface of theinert particles. Once the bound antigen / antibody are exposed to itsspecific antibody (usually IgM) or antigen, antigen-antibody complexeswill be formed.In the case of the particles with absorbed / bound antigens, due tothe presence of multiple Fab arms or the IgM, a single IgM may complexwith bound antigens from 2 adjacent particles. Meanwhile, in the case of the particles with absorbed / bound antibody, the soluble antigens (insample) that have multiple antigenic determinants will bind to antibodiesfrom 2 adjacent particles. In both cases, an antibody- antigen-particlenetwork is formed through multiple repeats of the aforementionedreactions. As a result, agglutination and clumping of the particlesoccurred.Indirect agglutination assay that uses erythrocytes as carrier of antigens is termed "Passive Haemagglutination" while assays usingantibody-coated erythrocytes are termed "Reverse PassiveHaemagglutination".
1 |Page
 
[PRACTICAL 7]MTEB 2404
RHEUMATOID FACTOR (RF) LATEX SLIDE TEST
PRINCIPLE
 The RF reagent is a suspension of polystyrene latex particlessensitized with specially prepared human IgG. The reagent is based onan immunological reaction between human IgG bound to biologicallyinert latex particles and rheumatoid factors in the test specimen.When serum containing rheumatoid factors is mixed with the latexreagent, visible agglutination occurs. The RF latex reagent sensitivityhas been adjusted to detect a minimum of 8 lU/mL of rheumatoidfactors according with the WHO International Standard without previoussample dilution.
REAGENTS
1.
RF Latex Reagent:
A suspension of uniform polystyrene particles coatedwith IgG (human) in glycine buffer, pH 8.2; reagent sensitivity isstandardized with the World Health Organization RF Standard.
MIXWELL BEFORE USING.
2.
RF Positive Control Serum:
A stabilized, prediluted human serumcontaining at least 8 lU/mL of RF.
3.
RF Negative Control Serum:
A stabilized, prediluted human serumcontaining less than 8 lU/mL of RF.
4.
Glycine-Saline Buffer (20x):
pH 8.2 ± O.IM glycine and 0.15M NaCI.
(Dilute buffer following instructions on the label before using.)
***Store all reagents at 2 - 8°C. DO NOT FREEZE.
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[PRACTICAL 7]MTEB 2404
RF Latex Reagent
Pipette/Stir Sticks
RF Positive ControL
 Timer
RF Negative Control
 Test Tubes
Glycine-Saline Buffer
Serological pipettes
Reaction Slide
PROCEDURE
Qualitative Test:
1.
Bring reagents and specimens to room temperature before use.
2.
Place one drop (50 ul) of the RF Positive Control on field #l of thereaction slide. Place one drop (50 ul) of the RF Negative Control on field#2. The remaining fields are used for test specimens.
3.
Using pipettes provided. Place one drop of the undiluted specimens onsuccessive fields. Retain Pipette/Stir Sticks for mixing step.
4.
Gently resuspend the RF Latex Reagent and add one drop to each testfield. Use pipette/ Stir Stick to spread reaction mixture over entire testfield.
5.
Rotate the slide manually or with a mechanical rotor at 80-100 rpm for2 minutes and read immediately under direct light.
3 |Page

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