重组M蛋白-乳胶凝集试验检测PRRS病毒血清抗体的研究

Development of a recombinant M protein-based latex agglutination test for detection of antibody against PRRS virus

利用纯化的 PRRS病毒重组 M蛋白致敏乳胶制成乳胶抗原 ,成功地建立了一种检测 PRRS病毒血清抗体的乳胶凝集试验 (L AT)诊断方法。用制备的乳胶 M抗原分别检测猪瘟、猪伪狂犬病、猪细小病毒病、猪弓形体病、猪衣原体病、猪乙型脑炎阳性血清 ,结果均为阴性 ,无交叉反应 ,说明建立的 L AT方法具有良好的特异性。用建立的乳胶凝集试验方法与国外IDEXX公司 PRRS病毒抗体检测试剂盒同时对 76份猪血清样本进行检测 ,结果表明建立的 L AT方法的特异性和敏感性均为 95 % ,两种方法的总符合率为 87% ,检出率基本一致。研究结果表明 L AT方法具有操作简便、快速、敏感性高、特异性强、价格低廉且可用于现场检测等优点 ,是一种适合基层兽医单位用于

Using the purified recombinant M protein of PRRSV as antigen,a latex agglutination test(LAT) was developed for detection of antibody against porcine reproductive and respiratory syndrome virus(PRRSV).The various factors and conditions for LAT were explored and determined.Latex sensitized with recombinant M protein showed no reaction with the antibodies against hog Cholera virus(HCV),porcine pseudorabies virus(PRV),Porcine Parvovirus(PPV),Toxoplasmosis and swine Chlamydia,and no agglutination reaction were also fund between PBS,negative serum of PRRSV with latex sensitized with recombinant M protein It was revealed that a recombinant M protein-based latex agglutination test had good specificity for detecting antibodies against PRRSV in serum.76 swine sera samples from several pig farms were detected by LAT and IDEXX PRRSV antibody Test Kit simultaneously.It was shown that both the specificity and sensitivity for the established LAT were 95%.The total coincidence rate of both methods reached 87%(67/76).The results indicated that the LAT was a simple,rapid,sensitive,specific,inexpensive method which is suitable for detecting antibody against PRRSV in serum and serological survey of PRRS.