Simultaneous detection of human norovirus GI, GII and SARS-CoV-2 by a quantitative one-step triplex RT-qPCR
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 revers...
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| Format: | Article |
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Frontiers Media S.A.
2024-01-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1269275/full |
| _version_ | 1797362375445184512 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Hua-Wei Yi |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-08T16:06:07Z |
| format | Article |
| id | doaj.art-161aa4f33e2e47eba2c9691b6a8d2b2e |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-03-08T16:06:07Z |
| publishDate | 2024-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-161aa4f33e2e47eba2c9691b6a8d2b2e2024-01-08T05:11:28ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2024-01-011410.3389/fmicb.2023.12692751269275Simultaneous detection of human norovirus GI, GII and SARS-CoV-2 by a quantitative one-step triplex RT-qPCRHua-Wei Yi0Hua-Wei Yi1Xian-Mo Wang2Xian-Mo Wang3Xin Tan4Cai-Zhi Ding5Chang-Li Zhang6Chang-Li Zhang7Jia-Hao Wu8Jia-Hao Wu9Qi Li10Qi Li11Chen-Qi Xin12Chen-Qi Xin13Wen Fan14Wen Fan15The First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, ChinaThe First People’s Hospital of Jingzhou, Jingzhou, Hubei, ChinaThe First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, ChinaThe First People’s Hospital of Jingzhou, Jingzhou, Hubei, ChinaHealth Science Center of Yangtze University, Jingzhou, Hubei, ChinaThe People’s Hospital of Songzi, Jingzhou, Hubei, ChinaThe First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, ChinaThe First People’s Hospital of Jingzhou, Jingzhou, Hubei, ChinaThe First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, ChinaThe First People’s Hospital of Jingzhou, Jingzhou, Hubei, ChinaThe First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, ChinaThe First People’s Hospital of Jingzhou, Jingzhou, Hubei, ChinaThe First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, ChinaThe First People’s Hospital of Jingzhou, Jingzhou, Hubei, ChinaThe First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, ChinaThe First People’s Hospital of Jingzhou, Jingzhou, Hubei, ChinaBackgroundThere 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.https://www.frontiersin.org/articles/10.3389/fmicb.2023.1269275/fullnorovirusSARS-CoV-2nucleic acid detectionRT-qPCRgenotyping |
| spellingShingle | 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 Simultaneous detection of human norovirus GI, GII and SARS-CoV-2 by a quantitative one-step triplex RT-qPCR Frontiers in Microbiology norovirus SARS-CoV-2 nucleic acid detection RT-qPCR genotyping |
| title | Simultaneous detection of human norovirus GI, GII and SARS-CoV-2 by a quantitative one-step triplex RT-qPCR |
| title_full | Simultaneous detection of human norovirus GI, GII and SARS-CoV-2 by a quantitative one-step triplex RT-qPCR |
| title_fullStr | Simultaneous detection of human norovirus GI, GII and SARS-CoV-2 by a quantitative one-step triplex RT-qPCR |
| title_full_unstemmed | Simultaneous detection of human norovirus GI, GII and SARS-CoV-2 by a quantitative one-step triplex RT-qPCR |
| title_short | Simultaneous detection of human norovirus GI, GII and SARS-CoV-2 by a quantitative one-step triplex RT-qPCR |
| title_sort | simultaneous detection of human norovirus gi gii and sars cov 2 by a quantitative one step triplex rt qpcr |
| topic | norovirus SARS-CoV-2 nucleic acid detection RT-qPCR genotyping |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1269275/full |
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