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利用PCR方法获得PRRSV M基因,使用原核表达系统p ET-28a(+)-BL21(DE3)诱导表达M蛋白。用表达的M蛋白建立检测PRRSV抗体的间接ELISA。通过特异性试验、阻断试验和重复性试验考查基于重组M蛋白建立的ELISA(M-ELISA)的实用性,同时使用M-ELISA和IDEXX Herd Chek ELISA for PRRSV试剂盒检测收集的400份临床血清样品。结果,用PCR扩增到了PRRSV M基因,获得了重组M蛋白,建立了用于检测PRRSV抗体的间接ELISA(M-ELISA);该M-ELISA具有良好的特异性和重复性。400份临床血清的检测结果与IDEXX试剂盒的检测结果的符合率为95.3%。上述结果表明,本研究基于表达的重组M蛋白,成功建立了检测RRSV抗体的间接ELISA。
The PRRSV M gene was obtained by PCR and the expression of M protein was induced by prokaryotic expression system p ET-28a (+) - BL21 (DE3). Indirect ELISA for detection of PRRSV antibodies was established using the expressed M protein. The utility of an ELISA (M-ELISA) established based on recombinant M protein was examined by specificity test, blocking test and repetitive test while the collected 400 serum samples were detected using M-ELISA and IDEXX Herd Chek ELISA for PRRSV kit sample. As a result, PRRSV M gene was amplified by PCR and recombinant M protein was obtained. An indirect ELISA (M-ELISA) for detecting PRRSV antibody was established; the M-ELISA has good specificity and repeatability. The coincidence rate of the detection results of 400 clinical serum and IDEXX kit was 95.3%. The above results indicate that this study established an indirect ELISA for the detection of RRSV antibodies based on the expressed recombinant M protein.