A one-step reverse-transcription recombinase aided PCR assay for the rapid and sensitive detection of human enteroviruses

Xiuli Sun; Huanhuan Lu; Yanqing Tie; Mengchuan Zhao; Ruiqing Zhang; Zhenlu Sun; Guohao Fan; Fengyu Li; Fengyu Tian; Yaxin Hu; Mengyi Zhang; Xinxin Shen; Xuejun Ma; Zhishan Feng

Biosafety and Health (Apr 2023)

A one-step reverse-transcription recombinase aided PCR assay for the rapid and sensitive detection of human enteroviruses

Xiuli Sun,
Huanhuan Lu,
Yanqing Tie,
Mengchuan Zhao,
Ruiqing Zhang,
Zhenlu Sun,
Guohao Fan,
Fengyu Li,
Fengyu Tian,
Yaxin Hu,
Mengyi Zhang,
Xinxin Shen,
Xuejun Ma,
Zhishan Feng

Affiliations

Xiuli Sun: North China University of Science and Technology, Tangshan 063210, China; Hebei General Hospital, Shijiazhuang 050051, China; NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
Huanhuan Lu: National Laboratory for Poliomyelitis, National Institute for Viral Disease Control and Prevention of the Chinese Center for Disease Control and Prevention, Beijing 102206, China
Yanqing Tie: Hebei General Hospital, Shijiazhuang 050051, China; Hebei Key Laboratory of Molecular Medicine, Shijiazhuang 050051, China; Hebei Clinical Research Center for Laboratory Medicine, Shijiazhuang 050051, China
Mengchuan Zhao: Department of Laboratory Medicine, Children's Hospital of Hebei Province, Shijiazhuang 050031, China
Ruiqing Zhang: NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
Zhenlu Sun: Yantai Center for Disease Control and Prevention, Yantai 264003, China
Guohao Fan: NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
Fengyu Li: Hebei General Hospital, Shijiazhuang 050051, China; NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
Fengyu Tian: Hebei General Hospital, Shijiazhuang 050051, China; NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
Yaxin Hu: North China University of Science and Technology, Tangshan 063210, China; Hebei General Hospital, Shijiazhuang 050051, China; NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
Mengyi Zhang: North China University of Science and Technology, Tangshan 063210, China; Hebei General Hospital, Shijiazhuang 050051, China; NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
Xinxin Shen: NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China; Corresponding authors: NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China (X. Shen, X. Ma); Hebei General Hospital, Shijiazhuang 050051, China (Z. Feng).
Xuejun Ma: NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China; Corresponding authors: NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China (X. Shen, X. Ma); Hebei General Hospital, Shijiazhuang 050051, China (Z. Feng).
Zhishan Feng: Hebei General Hospital, Shijiazhuang 050051, China; Hebei Key Laboratory of Molecular Medicine, Shijiazhuang 050051, China; Hebei Clinical Research Center for Laboratory Medicine, Shijiazhuang 050051, China; Corresponding authors: NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China (X. Shen, X. Ma); Hebei General Hospital, Shijiazhuang 050051, China (Z. Feng).

Journal volume & issue: Vol. 5, no. 2
pp. 126 – 131

Abstract

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Human enteroviruses (HEVs) include many different types that cause a wide range of diseases, and an effective method of genus-level identification has therefore significant clinical implications. However, quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), the gold-standard method, still has shortfalls in diagnostic sensitivity and timeliness. Here we established a one-step real-time reverse-transcription recombinase-aided PCR assay (RT-RAP) to detect HEV fragment within an hour. The RT-RAP assay showed a detection limit of 5 copies/μL using recombinant plasmids and was extensively verified using 15 HEV strains. Among 15 types of HEV (species A-C), the sensitivity of RT-RAP was approximately 2–8 folds lower than that of the qRT-PCR in 9 types, and no-cross reaction with other viruses was observed. RT-RAP was further applied to analyze CSF and fecal specimens; the clinical performance demonstrated that the RT-RAP and the commercial qRT-PCR kit provided consistent results. These results indicated that RT-RAP assay may be a promising approach for rapid and sensitive detection of HEV.

Published in Biosafety and Health

ISSN: 2590-0536 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Medicine: Internal medicine: Infectious and parasitic diseases; Medicine: Public aspects of medicine
Website: https://www.journals.elsevier.com/biosafety-and-health/

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