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

作　　者：李华玮[1] 付永[2] 黄岚 郭宏伟[1] 乔宏兴[3] 赵绪永[1] 边传周[3] LI Huawei;FU Yong;HUANG Lan;GUO Hongwei;QIAO Hongxing;ZHAO Xuyong;BIAN Chuanzhou(College of Food and Biological Engineering,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China;Academy of Animal Husbandry and Veterinary Sciences,Qinghai University,Xining 810016,China;School of Veterinary Medicine,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China)

机构地区：[1]河南牧业经济学院食品与生物工程学院,河南郑州450046 [2]青海大学畜牧兽医科学院,青海西宁810016 [3]河南牧业经济学院动物医药学院,河南郑州450046

出　　处：《云南农业大学学报（自然科学版）》2020年第1期62-67,共6页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家自然科学基金(31902284);河南省科技攻关项目(182102110021);河南牧业经济学院博士启动基金;河南牧业经济学院预防兽医重点学科(mxk2016102);河南牧业经济学院校科技创新团队(2018KYTD13)

摘　　要：【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.

关 键 词：猪繁殖与呼吸综合征病毒 免疫过氧化物酶单层细胞试验 检测 

分 类 号：S858.28[农业科学—临床兽医学]