牛副流感3型病毒Nano-PCR、LAMP方法的建立及初步应用  

Establishment and Preliminary Application of Nano-PCR and LAMP Methods for Bovine Parainfluenza Type-3 Virus

作　　者：李健友[1,2] 李家伟 王超 郭利[2] 何洪彬[1] 

机构地区：[1]东北农业大学,哈尔滨150030 [2]中国农业科学院特产研究所农业部经济动物疫病重点实验室,长春130122 [3]安图县畜牧兽医总站,安图133600

出　　处：《中国畜牧兽医》2017年第10期2837-2844,共8页China Animal Husbandry & Veterinary Medicine

基　　金：国家重点研发计划:畜禽重大疫病防控于高效安全养殖综合技术研发(2016YFD0500907);吉林省科技厅重点科技成果转化项目(20150307024NY);吉林省科技厅产业技术创新战略联盟项目(20170309002NY)

摘　　要：试验旨在建立牛副流感3型病毒(bovine parainfluenza type-3 virus,BPIV3)纳米PCR(Nano-PCR)与环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)新型快速分子检测技术,对其进行特异性和敏感性对比试验,并对10份临床样品进行了检测。特异性与敏感性试验结果显示,建立的Nano-PCR和LAMP方法只对BPIV3特异,而对牛呼吸道合胞体病毒、牛传染性鼻气管炎病毒、牛病毒性腹泻病毒无交叉反应;建立的Nano-PCR和LAMP方法具有相同的敏感性,均是普通PCR的10倍,最低核酸检出量均为4.16×102拷贝/μL。临床检测结果显示,建立的两种方法阳性符合率为100%,且阳性检出率均高于普通PCR。因此,本试验建立的Nano-PCR和LAMP方法为BPIV3的临床诊断提供了更快速、敏感、可靠的工具。This study was aimed to establish Nano-PCR and LAMP which were new rapid type of molecular detection technologies of bovine parainfluenza type-3 virus（ BPIV3）. The comparative specificity and sensitivity of BPIV3 Nano-PCR and LAMP PCR were tested,and the assay was applied to detect 10 clinical samples. The specificity and sensitivity tests showed that Nano-PCR and LAMP were only sensitive to BPIV3,without cross reaction to other viruses,such as bovine respiratory syncytial virus,infectious bovine rhinotracheitis virus and bovine viral diarrhea virus. In the sensitivity test,Nano-PCR and LAMP showed10 times sensitivity than that of the traditional PCR technology,with the minimum detection of 4. 16 ×102copies/μL. Clinical test results showed that the coincidence rate of Nano-PCR and LAMP could reach to100%,and the positive detection rate was much higher than that of normal PCR. Therefore,the NanoPCR and LAMP methods established in this study provided a faster,sensitive and reliable tool for the clinical diagnosis of BPIV3.

关 键 词：牛副流感3型病毒(BPIV3) Nano-PCR LAMP 

分 类 号：S858.23[农业科学—临床兽医学]