Rapid two-stage amplification in a single tube for simultaneous detection of norovirus GII and group a rotavirus.

Autor: Li FY; Hebei Medical University, Shijiazhuang, Hebei, China.; Hebei General Hospital, Shijiazhuang, Hebei, 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, China., Guo YH; Department of Clinical Laboratory, Children's Hospital of Hebei Province, Shijiazhuang, Hebei, China., Sun ZL; 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, China., Liu H; Shandong Provincial Research Center for Bioinformatic Engineering and Technique, School of Life Sciences and Medicine, Shandong University of Technology, Zibo, Shandong, China., Zhao MC; Department of Clinical Laboratory, Children's Hospital of Hebei Province, Shijiazhuang, Hebei, China., Cui J; Hebei General Hospital, Shijiazhuang, Hebei, China., Jiang Y; The Third Hospital of Hebei Medical University, Shijiazhuang City, Hebei Province, China., Shen XX; 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, China., Ma XJ; 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, China., Feng ZS; Hebei Medical University, Shijiazhuang, Hebei, China.; Hebei General Hospital, Shijiazhuang, Hebei, China.; Hebei Medical Laboratory Clinical Medical Research Center, Shijiazhuang City, Hebei Province, China.; Hebei Key Laboratory of Molecular Medicine, Shijiazhuang City, Hebei Province, China.
Jazyk: angličtina
Zdroj: Journal of clinical laboratory analysis [J Clin Lab Anal] 2023 Mar; Vol. 37 (5), pp. e24858. Date of Electronic Publication: 2023 Mar 14.
DOI: 10.1002/jcla.24858
Abstrakt: The most prevalent viruses currently causing diarrhea are norovirus and rotavirus, and rapid and sensitive detection methods are essential for the early diagnosis of disease. The purpose of this study was to establish a sensitive single-tube two-stage nucleic acid amplification method-reverse transcription recombinase-assisted PCR (RT-RAP)-for simultaneous detection of norovirus GII and group A Rotavirus, with the first stage consisting of isothermal reverse transcription recombinase-aided amplification (RT-RAA) and the second stage consisting of qPCR (quantitative PCR). RT-RAP is more sensitive than either RT-RAA or qRT-PCR (quantitative RT-PCR) alone. And the addition of a barrier that can be disassembled after heating enabled the detection of samples within 1 h in a single closed tube. Sensitivity was 10 copies/reaction of norovirus (Novs) GII and group A rotavirus (RVA). In parallel, two hundred fecal specimens were used to evaluate the method and compare it with a commercial fluorescent quantitative RT-PCR. The data showed kappa values of 0.957 and 0.98 (p < 0.05) for detecting Novs GII and RVA by the two methods, indicating the potential of the newly established assay to be applied to clinical and laboratory testing.
(© 2023 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC.)
Databáze: MEDLINE
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