猪繁殖与呼吸综合征病毒免疫过氧化物酶单层细胞检测方法的建立

Establishment of Immunoperoxidase Monolayer Assay for Porcine Reproductive and Respiratory Syndrome Virus

【目的】建立一种可直观判断猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)的检测方法。【方法】应用PRRSV多抗血清作为一抗,建立了PRRSV免疫过氧化物酶单层细胞检测体系,并对相关反应条件进行优化,对其特异性和敏感性进行检测。【结果】最佳病毒接种感染复数(multiplicity of infection,MOI)为0.1,最佳感染时间为24 h,一抗最佳稀释度为1∶1000,所建立的PRRSV免疫过氧化物酶单层细胞检测方法与其他几种流行猪病无交叉反应。对300份来自不同地区的临床血清进行检测,IPMA与PCR检测阳性符合率达93%。【结论】该方法具有良好的特异性和敏感性,为PRRSV的临床检测提供新方法。

[Purpose]In order to establish a method for visually determining porcine reproductive and respiratory syndrome virus detection.[Method]Anti-PRRSV polyclonal serum was used as the primary antibody in this study to establish a PRRSV immunoperoxidase monolayer assay system,and to optimize the relevant reaction conditions detecting its specificity and sensitivity.[Result]The best inoculation MOI value was 0.1 and the best infection time was 24 hours and the best dilution of primary antibody was 1:1000.[Conclusion]The established PRRSV immunoperoxidase monolayer assay had no cross-reaction with other endemic swine diseases.The positive coincidence rate of IPMA and PCR in 300 clinical serum samples collected from different regions reached 93%,indicating that this method has good specificity and sensitivity,which provides a new method for the clinical detection of PRRSV.