A recombinase polymerase amplification-based lateral flow strip assay for rapid detection of genogroup II noroviruses in
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ORIGINAL ARTICLE
A recombinase polymerase amplification‑based lateral flow strip assay for rapid detection of genogroup II noroviruses in the field Tianhui Jia1 · Yongxin Yu1 · Yongjie Wang1,2,3 Received: 25 February 2020 / Accepted: 4 August 2020 © Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract Human norovirus is the leading cause of viral gastroenteritis worldwide. Rapid detection facilitates management of disease outbreaks, but field diagnosis is difficult to achieve due to the lack of reliable and portable methods. Recombinase polymerase amplification (RPA) is a robust isothermal amplification method that is capable of rapidly amplifying and detecting nucleic acids using simple equipment. In this study, RPA combined with lateral flow (LF) strips specific for human genogroup II (GII) noroviruses was established and evaluated. The assay specifically detects purified GII noroviruses as well as RNA in boiled human stool samples, with a sensitivity of 50 norovirus genome copies per reaction. The whole detection procedure of the one-step RT-RPA-LF is completed within 20 min, which is eight times faster than that of the standard real-time RT-PCR. The RT-RPA-LF method described here is suitable for rapid field diagnosis of all GII noroviruses in human stool samples.
Introduction Norovirus is a genus of single-stranded, positive-sense, nonenveloped RNA viruses in the family Caliciviridae [1, 2]. Currently, noroviruses are divided into 10 genogroups (GIGX) and further divided into more than 49 genotypes [3]. Five genogroups of GI, GII, GIV, GVIII and GIX noroviruses infect humans [3]. Of these, GI and GII noroviruses are the most common ones, including at least 31 distinct genotypes [4]. Noroviruses are of particular concern in relatively closed environments, such as healthcare settings, schools, and kindergartens, and outbreaks cause high morbidity [5]. Early detection is important for monitoring outbreaks, Handling Editor: Reimar Johne. * Yongxin Yu [email protected] * Yongjie Wang [email protected] 1
College of Food Science and Technology, Shanghai Ocean University, Shanghai, China
2
Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China
3
Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai, China
which reduces health care costs and improves public health by facilitating more rapid implementation of strict exposure controls [6]. Molecular tests for noroviruses usually involve detection of viral nucleic acid by extraction of viral RNA, reverse transcription (RT), and amplification, which is carried out in a one- or two-step procedure [7], such as RT-PCR, nested RT-PCR, RT-quantitative PCR (qPCR), loop-mediated isothermal amplification (LAMP), or recombinase polymerase amplification (RPA) [8–11]. PCR-based molecular detection methods are typically used for detection of noroviruses. For example, nested RT-PCR can be used
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