Development Of Avian Leukosis Virus Antigen Test Kit

p27 gene is one of rather conservative genes among different subgroups of avian Leukosis virus. The capsid p27 protein is the major component of the core shell of avian avian Leukosis virus and the major group specific antigen. So p27 protein is the optimum antigen used to develop group specific antibody. In this study we used anti-p27 antibody to successfully develop an avian leucosis virus antigen test kit.Conditions were optimized for using these preparations for an ALV-specific ELISA. It was shown that the optimal concentration of anti-p27 McAb for coating of plate was 1.34μ g/ml, the blocking buffer was 5% skim milk, the optimal blocking condition for ELISA was 4℃, 2 hours, the sample should be incubated at 37℃ for 60 min, the working concentration of HRP-labeled rabbit anti-p27 antibody wasl:1200 and should be incubated at 37℃ for 60 min, the TMB substrate for ELISA was incubated at room temperature for 15 min, stop solution was 1M H₂SO₄, and the cut off value of ELISA was 0.17.The developed kit was used to detect E.coli , AIV ,NDV, EDS'76, IBDV etc. The results revealed that the ELISA had good specificity for the detection of ALV. We daily used this developed ELISA to test the same positive and negative samples for 4 days. There was no significant difference (P>0.05) among positive values or negative values, so the assay was demonstrate to have a good reproduction.276 albumin samples were tested for the ALV by using the developed ELISA and the IDEXX ALV antigen test kit simultaneously. It was found that the specificity of the developed ELISA was 100% compared with the IDEXX kit.The coated plates were used to test the same positive or negative sample for 6 times, one time per month. The OD values of positive sample were about 1.0 and the OD values of negative samples were lower than 0.1.The result demonstrated that the shelf-life of this kit was 6 six months. The study has developed an ELISA, which provides an available technique for diagnosing ALV and is a good base to further develop and merchandise the kit.