文献类型：期刊文献

中文题名：犬瘟热病毒巢式RT-PCR检测方法的建立及初步应用

英文题名：Development and Application of RT-nested PCR for Detecting Canine Distemper Virus

作者：杨井泉[1,2];韩猛立[2];朱艳平[3];黄新[2];薄新文[2]

第一作者：杨井泉

机构：[1]石河子大学动物科技学院;[2]新疆农垦科学院/新疆兵团绵羊繁育生物技术重点实验室;[3]新乡学院生命科学与技术系

第一机构：石河子大学动物科技学院,新疆石河子832003

年份：2011

卷号：48

期号：6

起止页码：1098-1103

中文期刊名：新疆农业科学

外文期刊名：Xinjiang Agricultural Sciences

收录：CSTPCD;;北大核心:【北大核心2008】；CSCD:【CSCD_E2011_2012】；

基金：国家产业技术体系专项(NYCYTX-40-13);家畜疫病病原生物学国家重点实验室开放基金课题(SKLVEB2009KFKT016)

语种：中文

中文关键词：犬瘟热病毒;反转录巢式PCR;lit—PCR;NP基因

外文关键词：canine distemper virus （CDV） ; RT-nested PCR; RT-PCR; NP gene

摘要：【目的】建立一种犬瘟热病毒(CDV)RT-nested PCR检测方法。【方法】根据CDV弱毒株Onderstepoort的核衣壳蛋白(NP)基因序列设计套式引物,进行特异性、敏感性、重复性实验。【结果】特异性试验表明,该方法可以特异扩增出CDV NP基因片段,但从RNA病毒狂犬病病毒(RV)、DNA病毒犬细小病毒(CPV)以及正常Vero细胞中均未扩增出该条带。敏感性试验表明,RT-PCR可以扩增10-3稀释度的病毒RNA,而建立的RT-nested PCR可以扩增10-6稀释度的病毒RNA,后者的敏感性明显高于前者。重复性实验表明,不同情况下的3次重复实验,结果相同。用RT-nested PCR对石河子地区疑似感染CDV的病料进行检测,结果表明,该研究建立的RT-nested PCR不仅能有效的检测CDV感染,而且能够检测不同组织的样品。【结论】建立的RT-nested PCR检测方法,适用于动物CDV的快速诊断和流行病学调查。
[ Objective] The chief purpose of this study was to establish ART- nested PCR assay for detecting Canine distemper virus （CDV）. [Method]The primers were designed by using nucleocap sid protein （NP） gene information of CDV Onderstepoort strain in GenBank. The repeatability, specificity and sensitivity of assay were evaluated. [Result] The specificity of assay suggested that the assay amplified only a NP gene from CDV： but not from other virus sueh as Rabies virus （ RV）, Canine parvovirus （CPV） and Vero cell. The sensitivity of assay suggested that serial 10 - fold dilutions of 10-3 CDV RNA were positive for RT- PCR, but the lower detection limit of CDV RNA 10-6 were positive for the RT - nested PCR. And the sensitivity of assay for RT - PCR was higher than that of RT - nested PCR. The repeated experiments for repeatability of assay were consistent. The RT - nested PCR method was successfully used to detect suspected CDV infection in clinical samples from Shihezi, and it might be useful for the clinical samples from different tissues. It also might be used for a quick diagnosis of CD and the epidemiological investigation. [ Conclusion ] The established RT- nested PCR method can be used for a quick diagnosis of CD and the epidemiological investigation.