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
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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DOI：
10.1016/j.jviromet.2016.04.012
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发表时间：
2016-08-01
影响因子：
3.1
通讯作者：
King, Donald P.
中科院分区：
文献类型：
--
作者：
Fowler, Veronica L.;Howson, Emma L. A.;King, Donald P.
水泡性口炎（VS）是中美洲和南美洲北方地区的地方病，在那里牛和猪中的零星爆发可引起与口蹄疫（FMD）相似的临床体征。因此，迫切需要快速，灵敏和特异性的鉴别诊断测定，适用于该领域的决策。已经开发了用于水泡性疾病如FMD和猪水泡病（SVD）的RT-LAMP测定，但是目前没有可以检测VS病毒（VSV）的RT-LAMP测定，也没有任何多重RT-LAMP测定允许原位快速区分这些“看起来像”的疾病。本研究描述了一种新的RT-LAMP检测VSV专注于新泽西（VSNJ）血清型，这导致了最近在美洲的VS案件的发展。该RT-LAMP测定以结合分子侧向流装置的多重形式组合，用于FMD和VS之间的区分。该测定能够检测代表性VSNJV，并且单重和多重VSNJV RT-LAMP测定的检测限等同于基于实验室的实时RT-PCR测定。开发了一种类似的多重RT-LAMP测定来区分FMDV和SVDV，表明FMDV、SVDV和VSNJV可以在上皮悬浮液中可靠地检测，而不需要预先提取RNA，提供了一种方法，该方法可以用作快速和低成本测定的基础，用于区分FMD与该领域的其他水泡性疾病。(C)2016年6月，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/）。
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/).