6种商品化PRRSV美洲经典毒株和高致病性毒株双重实时荧光定量PCR检测试剂盒性能比对  

Comparison of Six Commercial Duplex Real-time Fluorescence Quantitative PCR Kits for Classical American-type and Highly Pathogenic Strains of PRRSV

作　　者：高晓龙 栗云鹏 李志军 张启龙 傅彩霞 高敏 雷琪莉 王林 Gao Xiaolong;Li Yunpeng;Li Zhijun;Zhang Qilong;Fu Caixia;Gao Min;Lei Qili;Wang Lin(Beijing Center for Animal Disease Prevention and Control,Beijing 102629,China)

机构地区：[1]北京市动物疫病预防控制中心,北京102629

出　　处：《中国动物检疫》2025年第2期80-86,共7页China Animal Health Inspection

基　　金：北京市动物疫病预防控制中心综合保障项目(畜病监测)。

摘　　要：为评价不同品牌猪繁殖与呼吸综合征病毒(PRRSV)美洲经典毒株和高致病性毒株双重实时荧光定量PCR检测试剂盒的性能,选择6种市售商品化试剂盒,对其敏感性、特异性、重复性以及临床样品检测符合率等指标进行比对分析。结果显示:对于PRRSV美洲经典毒株VR2332和高致病性毒株JXA1,A品牌试剂盒最低检测限均可达到1 copy/μL,D品牌试剂盒最低检测限分别为100和1000 copies/μL,其他品牌试剂盒最低检测限为1~10 copies/μL;6种试剂盒对猪瘟病毒(CSFV)、伪狂犬病病毒(PRV)、猪流行性腹泻病毒(PEDV)、猪传染性胃肠炎病毒(TGEV)、猪细小病毒(PPV)的检测均为阴性,特异性良好;对不同浓度模板进行检测,除1个变异系数为6.5%以外,其余均小于5.0%,重复性较好。临床样品检测结果显示,F品牌试剂盒阴性样品检测符合率为95%,其余试剂盒为100%;D品牌试剂盒阳性样品检测符合率为50%,其余试剂盒为100%。综上所述,6种试剂盒的特异性、重复性均较好,但其敏感性及临床检测性能存在一定差异。建议在使用荧光定量PCR试剂盒检测PRRSV前,对其性能进行科学评估,选用性能良好的试剂盒以满足检测需求。本研究为检测机构和养殖场选择PRRSV检测试剂盒提供了依据。In order to evaluate the performance of duplex real-time fluorescence quantitative PCR kits with different brands for classical American-type porcine reproductive and respiratory syndrome virus(PRRSV)and highly pathogenic PRRSV,6 kinds of commercial kits were selected to compare and analyze their sensitivity,specificity and repeatability as well as the compliance rate of clinical sample detection.The results showed that,for classical American-type strain VR2332 and highly pathogenic strain JXA1,the lowest detection limits were both 1 copy/μL by kit A,which were 100 and 1000 copies/μL by kit D,respectively,and 1-10 copies/μL by other kits;6 kits failed to react with classical swine fever virus(CSFV),pseudorabies virus(PRV),porcine epidemic diarrhoea virus(PEDV),transmissible gastroenteritis virus(TGEV)and porcine parvovirus(PPV),presenting a good specificity;all coefficients of variation(except one)detected from the templates with different concentrations were less than 5%,indicating a good repeatability.The detection results of clinical samples showed that the compliance rate of negative samples was 95%detecting by kit F,and 100%by other kits;while that of positive samples was 50%by kit D,and 100%by other kits.In conclusion,all the 6 kits presented good specificity and repeatability,but different sensitivity and clinical detection performance to certain extent.It was recommended to scientifically evaluate the performance of the kits prior to PRRSV detection,and to select those with good performance to meet the needs of detection.Testing institutions and farms were provided with a basis to select appropriate kits for PRRSV.

关 键 词：猪繁殖与呼吸综合征病毒 美洲经典毒株 高致病性毒株 PCR检测试剂盒 性能对比 

分 类 号：S851.4[农业科学—预防兽医学]