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

Abstract：

: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 °C for 40 min. Specificity of 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.

journal_name

J Virol Methods

journal_title

Journal of virological methods

authors

Nemoto M,Morita Y,Niwa H,Bannai H,Tsujimura K,Yamanaka T,Kondo T

doi

10.1016/j.jviromet.2015.02.001

subject

Has Abstract

pub_date

2015-04-01 00:00:00

pages

13-6

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(15)00023-3

journal_volume

215-216

pub_type

杂志文章

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