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采用多重PCR和PCR-RFLP技术对本地治里地区犬细小病毒进行分子分型

Molecular Typing of Canine parvovirus Occurring in Pondicherry by Multiplex PCR and PCR-RFLP.

作者信息

Parthiban S, Mukhopadhyay H K, Antony P X, Pillai R M

机构信息

Rajiv Gandhi College of Veterinary and Animal Sciences, 10/442, 1st Cross, Thulukathamman Nagar, Murungapakkam Post, Pondicherry, 605004 India.

出版信息

Indian J Virol. 2010 Jun;21(1):86-9. doi: 10.1007/s13337-010-0011-2. Epub 2010 Aug 19.

Abstract

The present study was aimed at molecular typing of Canine parvovirus (CPV) occurring in Pondicherry using PCR based assays. CPV-2a and CPV-2b types were detected by PCR in 68 (53.12%) out of 128 faecal samples/rectal swabs. All the 68 samples found positive by PCR assay were subjected to multiplex PCR assay with CPV-2ab and CPV-2b primer pairs. Sixty-seven (98.52%) samples were characterized as CPV-2b type and one sample (1.47%) was categorized as CPV-2a type. Sixty clinical samples found negative by CPV-2ab primers were subjected to another PCR assay with CPV-555 primer pair for detecting CPV-2c. Though three samples (5%) responded to this PCR, subsequent RFLP of these PCR products with MboII did not show any cleavage indicating the absence of CPV-2c in Pondicherry. It was inferred that CPV-2b was the most prevalent CPV type in Pondicherry. It was further concluded that the CPV-2 variants (CPV-2a, CPV-2b and CPV-2c) currently circulating in the field worldwide could be diagnosed by employing multiplex PCR and PCR-RFLP assays.

摘要

本研究旨在利用基于聚合酶链反应(PCR)的检测方法,对本地治里地区出现的犬细小病毒(CPV)进行分子分型。在128份粪便样本/直肠拭子中,通过PCR检测到68份(53.12%)含有CPV-2a和CPV-2b型。所有经PCR检测呈阳性的68份样本,均使用CPV-2ab和CPV-2b引物对进行多重PCR检测。其中67份(98.52%)样本被鉴定为CPV-2b型,1份样本(1.47%)被归类为CPV-2a型。60份经CPV-2ab引物检测呈阴性的临床样本,使用CPV-555引物对进行另一项PCR检测,以检测CPV-2c。尽管有3份样本(5%)对该PCR有反应,但随后用MboII对这些PCR产物进行限制性片段长度多态性分析(RFLP)时,未显示任何切割,表明本地治里地区不存在CPV-2c。由此推断,CPV-2b是本地治里地区最普遍的CPV类型。进一步得出结论,目前在全球范围内流行的CPV-2变异株(CPV-2a、CPV-2b和CPV-2c)可通过多重PCR和PCR-RFLP检测进行诊断。

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