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首页> 外文期刊>Journal of Virological Methods >Development of a reverse transcription loop-mediated isothermal amplification assay for the detection of vesicular stomatitis New Jersey virus: Use of rapid molecular assays to differentiate between vesicular disease viruses
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Development of a reverse transcription loop-mediated isothermal amplification assay for the detection of vesicular stomatitis New Jersey virus: Use of rapid molecular assays to differentiate between vesicular disease viruses

机译：逆转录环介导的等温扩增测定法的开发，用于检测水疱性口炎新泽西州病毒：使用快速分子测定法区分水疱性疾病病毒

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摘要

Vesicular stomatitis (VS) is endemic in Central America and northern regions of South America, where sporadic outbreaks in cattle and pigs can cause clinical signs that are similar to foot-and-mouth disease (FMD). There is therefore a pressing need for rapid, sensitive and specific differential diagnostic assays that are suitable for decision making in the field. RT-LAMP assays have been developed for vesicular diseases such as FMD and swine vesicular disease (SVD) but there is currently no RT-LAMP assay that can detect VS virus (VSV), nor are there any multiplex RT-LAMP assays which permit rapid discrimination between these 'look-a-like' diseases in situ. This study describes the development of a novel RT-LAMP assay for the detection of VSV focusing on the New Jersey (VSNJ) serotype, which has caused most of the recent VS cases in the Americas. This RT-LAMP assay was combined in a multiplex format combining molecular lateral-flow devices for the discrimination between FMD and VS. This assay was able to detect representative VSNJV's and the limit of detection of the singleplex and multiplex VSNJV RT-LAMP assays were equivalent to laboratory based real-time RT-PCR assays. A similar multiplex RT-LAMP assay was developed to discriminate between FMDV and SVDV, showing that FMDV, SVDV and VSNJV could be reliably detected within epithelial suspensions without the need for prior RNA extraction, providing an approach that could be used as the basis for a rapid and low cost assay for differentiation of FMD from other vesicular diseases in the field. (C) 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.orgilicenses/by/4.0/).
机译：水泡性口炎（VS）在中美洲和南美洲北部地区是地方性流行，在那里牛和猪的零星爆发会导致类似于口蹄疫（FMD）的临床症状。因此，迫切需要适合于现场决策的快速，灵敏和特异的鉴别诊断测定。已经针对诸如FMD和猪水泡病（SVD）的水疱疾病开发了RT-LAMP测定法，但是目前没有可以检测VS病毒（VSV）的RT-LAMP测定法，也没有任何可以快速检测的多重RT-LAMP测定法这些“相似”疾病之间的区别。这项研究描述了一种新的RT-LAMP分析方法的开发，该方法主要用于检测新泽西州（VSNJ）血清型的VSV，这已导致美洲最近发生的大多数VS病例。将该RT-LAMP测定法以多种形式组合，结合分子横向流动装置以区分FMD和VS。该测定法能够检测代表性的VSNJV，单重和多重VSNJV RT-LAMP测定法的检出限等同于基于实验室的实时RT-PCR测定法。开发了一种类似的多重RT-LAMP测定法来区分FMDV和SVDV，这表明可以在上皮悬浮液中可靠地检测FMDV，SVDV和VSNJV，而无需事先进行RNA提取，从而提供了一种可以用作基础的方法快速，低成本的检测方法，用于将FMD与本领域其他水泡疾病区分开来。 （C）2016作者。由Elsevier B.V.发布。这是CC BY许可下的开放获取文章（http：//creativecommons.orgilicenses/by/4.0/）。

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