通过定量一步三重RT-qPCR同时检测人诺如病毒GI、GII和严重急性呼吸综合征冠状病毒2
Simultaneous detection of human norovirus GI, GII and SARS-CoV-2 by a quantitative one-step triplex RT-qPCR.
作者信息
Yi Hua-Wei, Wang Xian-Mo, Tan Xin, Ding Cai-Zhi, Zhang Chang-Li, Wu Jia-Hao, Li Qi, Xin Chen-Qi, Fan Wen
机构信息
The First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, China.
The First People's Hospital of Jingzhou, Jingzhou, Hubei, China.
出版信息
Front Microbiol. 2024 Jan 8;14:1269275. doi: 10.3389/fmicb.2023.1269275. eCollection 2023.
BACKGROUND
There are many similarities in the clinical manifestations of human norovirus and SARS-CoV-2 infections, and nucleic acid detection is the gold standard for diagnosing both diseases. In order to expedite the identification of norovirus and SARS-CoV-2, a quantitative one-step triplex reverse transcription PCR (RT-qPCR) method was designed in this paper.
METHODS
A one-step triplex RT-qPCR assay was developed for simultaneous detection and differentiation of human norovirus GI (NoV-GI), GII (NoV-GII) and SARS-CoV-2 from fecal specimens.
RESULTS
The triplex RT-qPCR assay had high detection reproducibility (CV < 1%) and sensitivity. The lower limits of detection (LLOD95) of the triplex RT-qPCR assay for each target site were 128.5-172.8 copies/mL, and LLOD95 of the singleplex RT-qPCR assay were 110.3-142.0 copies/mL. Meanwhile, among the detection of clinical oropharyngeal swabs and fecal specimens, the results of the singleplex and triplex RT-qPCR assay showed high agreement.
CONCLUSION
The triplex RT-qPCR assay for simultaneous detection of NoV-GI, NoV-GII and SARS-CoV-2 from fecal specimens has high clinical application value.
背景
人类诺如病毒感染与新型冠状病毒肺炎（SARS-CoV-2）感染在临床表现上有许多相似之处，核酸检测是这两种疾病诊断的金标准。为加快诺如病毒和SARS-CoV-2的鉴定，本文设计了一种定量一步三重逆转录聚合酶链反应（RT-qPCR）方法。
方法
建立了一种一步三重RT-qPCR检测方法，用于同时从粪便标本中检测和区分人类诺如病毒GI型（NoV-GI）、GII型（NoV-GII）和SARS-CoV-2。
结果
三重RT-qPCR检测具有较高的检测重复性（CV<1%）和灵敏度。三重RT-qPCR检测各靶点的最低检测限（LLOD95）为128.5-172.8拷贝/毫升，单重RT-qPCR检测的LLOD95为110.3-142.0拷贝/毫升。同时，在临床咽拭子和粪便标本检测中，单重和三重RT-qPCR检测结果高度一致。
结论
粪便标本同时检测NoV-GI、NoV-GII和SARS-CoV-2的三重RT-qPCR检测方法具有较高的临床应用价值。
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