Haemagglutination Assay
HAEMAGGLUTINATION TEST
■ Haemagglutinin proteins (H-16) has the ability to agglutinate the Red Blood Cell which is an indicator of the presence of virus.
■ Magnitude of the agglutination depends on quantity of virus.
■ Two fold serial dilution of the suspected virus material in P.B.S is treated with 1% washed R.B.C.s.
■ Virus agglutination can be seen in micro titration plates.
■ Reciprocal of the last virus dilution giving agglutination is Haemagglutination titer (HA titer).
[pic]
HAEMAGGLUTINATION INHIBITION TEST
■ Haemagglutination activity of the virus can be inhibited by the addition of the specific antibodies present in serum or specific antisera.
■ Field serum samples are serially diluted in P.B.S & allowed to react with standard antigens
(4 HA).
■ Half an hour is given to the reaction so that antigen and antibody may react.
■ 1% R.B.Cs are added to evaluate the Haemagglutination inhibition.
■ Serum samples containing specific antibodies against the antigen used will inhibit the haemagglutination activity of the virus.
■ Reciprocal of the last serum dilution causing Haemagglutination inhibition will consider as (HI titer).
■ Positive serum control, negative serum control, antigen control, R.B.Cs control are run to validate the test result.
[pic]
ENZYME LINKED IMMUNOSORBANT ASSAY
(ELISA)
■ PRINCIPLE
■ Antigen and antibody reaction are highly specific.
■ Antibody’s FC portion can be attached with enzyme.
■ Specific antigen & antibody reactions with enzyme can be detected by the addition of the substrate specific to the enzyme.
■ Magnitude of antigen and antibody present in the reaction seen with the intensity of colors cab be read by ELISA reader.
[pic]
Types of ELISA
■ Direct ELISA
■ Indirect ELISA
■ Sandwich ELISA
■ Competitive ELISA
■ Multiplex ELISA
[pic]
AGAR GEL
■ Haemagglutinin proteins (H-16) has the ability to agglutinate the Red Blood Cell which is an indicator of the presence of virus.
■ Magnitude of the agglutination depends on quantity of virus.
■ Two fold serial dilution of the suspected virus material in P.B.S is treated with 1% washed R.B.C.s.
■ Virus agglutination can be seen in micro titration plates.
■ Reciprocal of the last virus dilution giving agglutination is Haemagglutination titer (HA titer).
[pic]
HAEMAGGLUTINATION INHIBITION TEST
■ Haemagglutination activity of the virus can be inhibited by the addition of the specific antibodies present in serum or specific antisera.
■ Field serum samples are serially diluted in P.B.S & allowed to react with standard antigens
(4 HA).
■ Half an hour is given to the reaction so that antigen and antibody may react.
■ 1% R.B.Cs are added to evaluate the Haemagglutination inhibition.
■ Serum samples containing specific antibodies against the antigen used will inhibit the haemagglutination activity of the virus.
■ Reciprocal of the last serum dilution causing Haemagglutination inhibition will consider as (HI titer).
■ Positive serum control, negative serum control, antigen control, R.B.Cs control are run to validate the test result.
[pic]
ENZYME LINKED IMMUNOSORBANT ASSAY
(ELISA)
■ PRINCIPLE
■ Antigen and antibody reaction are highly specific.
■ Antibody’s FC portion can be attached with enzyme.
■ Specific antigen & antibody reactions with enzyme can be detected by the addition of the substrate specific to the enzyme.
■ Magnitude of antigen and antibody present in the reaction seen with the intensity of colors cab be read by ELISA reader.
[pic]
Types of ELISA
■ Direct ELISA
■ Indirect ELISA
■ Sandwich ELISA
■ Competitive ELISA
■ Multiplex ELISA
[pic]
AGAR GEL