title: Development of a minor groove binder assay for real-time PCR detection of porcine Sapelovirus.
abstract: A 5' 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' untranslated region (UTR) gene were designed. The assay was capable of detecting about 103copies/μl 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.
authors.affiliation: State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, China.
authors.forename: Junwei
authors.initials: J
authors.lastname: Chen
authors.affiliation: College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Chen
authors.affiliation: Guangdong Wen's Group Academy, Guangdong Wen's Foodstuffs Group Co., Ltd., Xinxing, Guangdong, China.
authors.forename: Qingfeng
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Guangdong Wen's Group Academy, Guangdong Wen's Foodstuffs Group Co., Ltd., Xinxing, Guangdong, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: Guangdong Wen's Group Academy, Guangdong Wen's Foodstuffs Group Co., Ltd., Xinxing, Guangdong, China.
authors.forename: Yanshan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Guangdong Wen's Group Academy, Guangdong Wen's Foodstuffs Group Co., Ltd., Xinxing, Guangdong, China.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Song
authors.affiliation: College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Xiangbin
authors.initials: X
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, China.
authors.forename: Chunyi
authors.initials: C
authors.lastname: Xue
authors.affiliation: College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Yingzuo
authors.initials: Y
authors.lastname: Bi
authors.affiliation: State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, China. Electronic address: caoych@mail.sysu.edu.cn.
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Cao
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D000818:Animals; D017931:DNA Primers / Q000235:genetics; D012328:RNA Viruses / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000821:virology*
chemical_list: D020121:5' Untranslated Regions; D017931:DNA Primers; D012367:RNA, Viral
keywords: Detection; Minor groove binder probe; Porcine Sapelovirus; Real-time PCR