Question

Now that the anti-CRP antibody has been shown to be pure and specific for CRP, the PI would like to look for the presence of CRP in the gall bladder. What test (ELISA, immunoblot, etc.) would you use and why?

Answer 1

CRP ( C- reactive protein) test is a blood test designed to measure the amount of CRP in the blood. C- reactive protein is an acute phase reactant, a protein made by liver and released into the blood in response to inflammation, tissue injury or infections.

AGGLUTINATION test is used for testing presence of CRP in gall ballder.

Principle : The C- reactive test is based on Rapid latex Agglutination Test. The test is based on the reaction between patients serum containing CRP as the antigen and corresponding antibody coated to the treated surface of latex particle. The coated particles enhance the detection of an agglutinate reaction when antigen is present in the serum being tested.

Latex Agglutination Test is used for CRP testing because the latex coated antibody react with antigen present in the serum to give visible Agglutination within two minutes.

In qualitative test, the reagents and sample are brought to room temperature. Place one drop of Serum from patient, postive control and negative control. Then add one drop of CRP latex reagent to each of the circles . Mix with separate mixing sticks and spread the fluid over the entire area of the cell. Tilt the slide back and forth slowly for two minutes and observe under artificial light. Observe for visible Agglutination.

Postive : visible Agglutination seen

Negative : no Agglutination seen

Calculation titre

CRP ug/ ml = 7× D, where D is the highest dilution of serum showing Agglutination and 7 is the sensitivity factor in ug/ ml.