| A silver-enhanced colloidal gold assay (SECGA) was developed for the detection of pseudo -rabies virus(PRV) antigen for the first time. The optimum working concentration of rabbit anti-PRV immunoglobulin G(IgG) was determined to be 1:400, that of sheep anti-rabbit IgG labeled with 5nm colloidal gold to be 1:40 and the optimum time of silver staining to be 15 minutes.The detection limit of purified PRV antigen by the SECGA was O.f5625 ug/ml, with the sensitivity of 8-fold greater than the indirect dot enzyme-linked immunos -orbent assay(DOT-ELISA). The high specity of the SECGA was show by the specific blocking test of rabbit anti-PRV serum and cross-reaction test with Porcine parvovirus(PPV), Suis fever virus(SFV), Pasteurella multocida and nomal cell culture.The positive rate of PRV of the 30 experimentally55infected rabbit specimens measured by the SECGA and indirect DOT-ELISA were 100%, while the positive rate of PRV of the 20 field specimens were 65% and 60%. The agreement rate of PRV detection by the SECGA and indirect DOT-ELISA was 97.8% in all the specimens, showing a high accordance between PRV antigen measured by the two tests.This study indicates that the SECGA is a low-cost, facile, rapid, highly sensitive, highly specific and reproducible method for the detection of PRV antigen. It is an efficient means for rapid diagnosis and epidemiological investigation of PRV infection. The study will lay a foundation for the application of SECGA in diagnoses and studies of other diseases in future. |
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