Development of a minor groove binder assay for real-time PCR detection of porcine Sapelovirus

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• 作者：Junwei Chen ; Feng Chen ; Qingfeng Zhou ; Wei Li ; Yanshan Chen ; Yanhua Song ; Xiangbin Zhang ; Chunyi Xue ; Yingzuo Bi ; Yongchang Cao
• 关键词：Porcine Sapelovirus ; Real-time PCR ; Minor groove binder probe ; Detection
• 刊名：Journal of Virological Methods
• 出版年：15 March, 2014
• 年：2014
• 卷：198
• 期：Complete
• 页码：69-74
• 全文大小：936 K

文摘

A 5鈥?conjugated minor groove binder (MGB) probe real-time PCR assay was developed in this study for porcine sapelovirus (PSV) detection and quantitation. Two primers and a MGB probe for the 5鈥?untranslated region (UTR) gene were designed. The assay was capable of detecting about 103 copies/渭l of standard template per reaction. Moreover, it does not detect any of the other RNA viruses that cause diarrhea disease in pigs. The coefficients of variation of intra- and inter-assay reproducibility were both lower than 2%. In 73 field fecal samples, PSV was detected in 46 samples using real-time PCR assay and only 32 samples with a conventional PCR assay. Therefore, the availability of this assay will facilitate further studies on the epidemiology of PSV infection and its role in swine disease.