論文

査読有り
2015年4月

Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification

JOURNAL OF VIROLOGICAL METHODS
  • Manabu Nemoto
  • ,
  • Yoshinori Morita
  • ,
  • Hidekazu Niwa
  • ,
  • Hiroshi Bannai
  • ,
  • Koji Tsujimura
  • ,
  • Takashi Yamanaka
  • ,
  • Takashi Kondo

215
開始ページ
13
終了ページ
16
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1016/j.jviromet.2015.02.001
出版者・発行元
ELSEVIER SCIENCE BV

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the rapid detection of equine coronavirus (ECoV). This assay was conducted at 60 degrees C for 40 min. Specificity the RT-LAMP assay was confirmed using several equine intestinal and respiratory pathogens in addition to ECoV. The novel assay failed to cross-react with the other pathogens tested, suggesting it is highly specific for ECoV. Using artificially synthesized ECoV RNA, the 50% detection limit of the RT-LAMP assay was 10(1.8) copies/reaction. This is a 50-fold greater sensitivity than conventional reverse transcription polymerase chain reaction (RT-PCR) assays, but a 4-fold lower sensitivity than quantitative RT-PCR (qRT-PCR) assays. Eighty-two fecal samples collected during ECoV outbreaks were analyzed. ECoV was detected in 59 samples using the RT-LAMP assay, and in 30 and 65 samples using RT-PCR or qRT-PCR assays, respectively. Although the RT-LAMP assay is less sensitive than qRT-PCR techniques, it can be performed without the need for expensive equipment. Thus, the RT-LAMP assay might be suitable for large-scale surveillance and diagnosis of ECoV infection in laboratories with limited resources. (C) 2015 Elsevier B.V. All rights reserved.

リンク情報
DOI
https://doi.org/10.1016/j.jviromet.2015.02.001
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/25682750
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000352329900003&DestApp=WOS_CPL
ID情報
  • DOI : 10.1016/j.jviromet.2015.02.001
  • ISSN : 0166-0934
  • eISSN : 1879-0984
  • PubMed ID : 25682750
  • Web of Science ID : WOS:000352329900003

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