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Defining the relative performance of isothermal assays that can be used for rapid and sensitive detection of foot-and-mouth disease virus

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タイトル: Defining the relative performance of isothermal assays that can be used for rapid and sensitive detection of foot-and-mouth disease virus
著者: Howson, Emma L.A. / Kurosaki, Yohei / Yasuda, Jiro / Takahashi, Masayoshi / Goto, Hiroaki / Gray, Ashley R. / Mioulet, Valerie / King, Donald P. / Fowler, Veronica L.
発行日: 2017年11月
出版者: Elsevier B.V.
引用: Journal of Virological Methods, 249, pp.102-110; 2017
抄録: This study describes the first multiway comparison of portable isothermal assays for the detection of foot-and-mouth disease virus (FMDV), benchmarked against real-time reverse transcription RT-PCR (rRT-PCR). The selected isothermal chemistries included reverse transcription loop-mediated isothermal amplification (RT-LAMP) and reverse transcription recombinase polymerase amplification (RT-RPA). The analytical sensitivity of RT-LAMP was comparable to rRT-PCR (101 RNA copies), while RT-RPA was one log10 less sensitive (102 RNA copies). Diagnostic performance was evaluated using a panel of 35 samples from FMDV-positive cattle and eight samples from cattle infected with other vesicular viruses. Assay concordance for RT-LAMP and RT-RPA was 86–98% and 67–77%, respectively, when compared to rRT-PCR, with discordant samples consistently having high rRT-PCR cycle threshold values (no false-positives were detected for any assay). In addition, a hierarchy of sample preparation methods, from robotic extraction to simple dilution of samples, for epithelial suspensions, serum and oesophageal-pharyngeal (OP) fluid were evaluated. Results obtained for RT-LAMP confirmed that FMDV RNA can be detected in the absence of RNA extraction. However, simple sample preparation methods were less encouraging for RT-RPA, with accurate results only obtained when using RNA extraction. Although the evaluation of assay performance is specific to the conditions tested in this study, the compatibility of RT-LAMP chemistry with multiple sample types, both in the presence and absence of nucleic acid extraction, provides advantages over alternative isothermal chemistries and alternative pen-side diagnostics such as antigen-detection lateral-flow devices. These characteristics of RT-LAMP enable the assay to be performed over a large diagnostic detection window, providing a realistic means to rapidly confirm positive FMD cases close to the point of sampling.
キーワード: Foot-and-Mouth disease / FMDV / Diagnostics / rRT-PCR / RT-LAMP / RT-RPA
URI: http://hdl.handle.net/10069/37782
ISSN: 01660934
DOI: 10.1016/j.jviromet.2017.08.013
権利: © 2017 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/).
資料タイプ: Journal Article
原稿種類: publisher
出現コレクション:120 学術雑誌論文

引用URI : http://hdl.handle.net/10069/37782



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