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

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

JOURNAL=Frontiers in Microbiology

VOLUME=14

YEAR=2024

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2023.1269275

DOI=10.3389/fmicb.2023.1269275

ISSN=1664-302X

ABSTRACT=<sec id="sec1"><title>Background</title><p>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.</p></sec><sec id="sec2"><title>Methods</title><p>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.</p></sec><sec id="sec3"><title>Results</title><p>The triplex RT-qPCR assay had high detection reproducibility (CV &lt; 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.</p></sec><sec id="sec4"><title>Conclusion</title><p>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.</p></sec>