Xin Tan

Yangtze University

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Changli Zhang

Nanjing Xiaozhuang University

Wen Fan

First People's Hospital of Jingzhou

Cai-Zhi Ding

Hua-Wei Yi

First People's Hospital of Jingzhou

Qi Li

Chinese Academy of Social Sciences

Xian-Mo Wang

First People's Hospital of Jingzhou

Chenqi Xin

Yangtze University

Jiahao Wu

Sichuan University

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Yangtze University

First People's Hospital of Jingzhou

Jilin University

Nanjing Medical University

Kunming Medical University

Wuhan University

Nanjing Xiaozhuang University

University of Science and Technology Beijing

Royal Society of Chemistry

Centre National pour la Recherche Scientifique et Technique (CNRST)

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Simultaneous Detection of Human Norovirus Gi, Gii and Sars-Cov-2 by a Quantitative One-Step Triplex Rt-Qpcr

Hua-Wei Yi Xian-Mo Wang Xin Tan Cai-Zhi Ding Changli Zhang

Background: The clinical manifestations of norovirus and SARS-CoV-2 infections are significantly similar, 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 N gene of SARS-CoV-2.ResultsThe triplex RT-qPCR assay had high detection reproducibility (CV<1%) and sensitivity. The lower limits of detection (LLOD95) of singleplex and triplex RT-qPCR assay for each target site were 110.1-157.1 copies/mL and 153.2-197.6 copies/mL, respectively. Meanwhile, among the detection of clinical oropharyngeal swabs and fecal specimens, the results of the singleplex and triplex RT-qPCR assay showed high agreement.Conclusions: The triplex RT-qPCR assay for simultaneous detection of NoV-GI, NoV-GII and SARS-CoV-2 has high clinical application value.

Sars virus

2019-20 coronavirus outbreak

10.2139/ssrn.4450383

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Simultaneous detection of human norovirus GI, GII and SARS-CoV-2 by a quantitative one-step triplex RT-qPCR

Frontiers in Microbiology (2024)

Hua-Wei Yi Xian-Mo Wang Xin Tan Cai-Zhi Ding Changli Zhang

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.

2019-20 coronavirus outbreak

Sars virus

10.3389/fmicb.2023.1269275

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