Frontiers in Microbiology (Jan 2024)

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

  • Hua-Wei Yi,
  • Hua-Wei Yi,
  • Xian-Mo Wang,
  • Xian-Mo Wang,
  • Xin Tan,
  • Cai-Zhi Ding,
  • Chang-Li Zhang,
  • Chang-Li Zhang,
  • Jia-Hao Wu,
  • Jia-Hao Wu,
  • Qi Li,
  • Qi Li,
  • Chen-Qi Xin,
  • Chen-Qi Xin,
  • Wen Fan,
  • Wen Fan

DOI
https://doi.org/10.3389/fmicb.2023.1269275
Journal volume & issue
Vol. 14

Abstract

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BackgroundThere 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.MethodsA 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.ResultsThe 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.ConclusionThe triplex RT-qPCR assay for simultaneous detection of NoV-GI, NoV-GII and SARS-CoV-2 from fecal specimens has high clinical application value.

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