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

机译:多重PCR和PCR-RFLP技术对本地治里犬细小病毒的分子分型

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摘要

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.
机译:本研究的目标是使用基于PCR的检测方法对本地治里犬细小病毒(CPV)进行分子分型。通过PCR在128份粪便样本/直肠拭子中检测到CPV-2a和CPV-2b类型(53.12%)。通过PCR测定发现阳性的所有68个样品均使用CPV-2ab和CPV-2b引物对进行多重PCR测定。 67个样本(98.52%)被表征为CPV-2b类型,一个样本(1.47%)被归类为CPV-2a类型。用CPV-555引物对对60个被CPV-2ab引物阴性的临床样品进行另一轮PCR检测,以检测CPV-2c。尽管三个样品(5%)对此PCR反应,但随后这些PCR产物用MboII的RFLP未显示任何裂解,表明本地治里没有CPV-2c。据推断,CPV-2b是本地治里最流行的CPV类型。进一步得出的结论是,可以通过使用多重PCR和PCR-RFLP分析来诊断目前在世界范围内流通的CPV-2变体(CPV-2a,CPV-2b和CPV-2c)。

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