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; Chang-Li Zhang; Jia-Hao Wu; Qi Li; Chen-Qi Xin; Wen Fan

doi:10.3389/fmicb.2023.1269275

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

Front Microbiol. 2024 Jan 8:14:1269275. doi: 10.3389/fmicb.2023.1269275. eCollection 2023.

Authors

Hua-Wei Yi^#^{1

2}, Xian-Mo Wang^#^{1

2}, Xin Tan³, Cai-Zhi Ding⁴, Chang-Li Zhang^{1

2}, Jia-Hao Wu^{1

2}, Qi Li^{1

2}, Chen-Qi Xin^{1

2}, Wen Fan^{1

2}

Affiliations

¹ The First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, China.
² The First People's Hospital of Jingzhou, Jingzhou, Hubei, China.
³ Health Science Center of Yangtze University, Jingzhou, Hubei, China.
⁴ The People's Hospital of Songzi, Jingzhou, Hubei, China.

^# Contributed equally.

Abstract

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.

Keywords: RT-qPCR; SARS-CoV-2; genotyping; norovirus; nucleic acid detection.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the Natural Science Foundation of Hubei Province, China [grant number 2021CFB261 (to H-WY)], Jingzhou Science and Technology Plan Projects, Hubei Province, China [grant number 2023HC82 (to H-WY) and 2023HC31 (to C-QX)] and the Doctoral Research Start-up Fund Project of the First People’s Hospital of Jingzhou, China [grant number 2023DIF03 (to H-WY)].