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

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

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

Afiliação

Yi HW; The First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, China.
Wang XM; The First People's Hospital of Jingzhou, Jingzhou, Hubei, China.
Tan X; The First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, China.
Ding CZ; The First People's Hospital of Jingzhou, Jingzhou, Hubei, China.
Zhang CL; Health Science Center of Yangtze University, Jingzhou, Hubei, China.
Wu JH; The People's Hospital of Songzi, Jingzhou, Hubei, China.
Li Q; The First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, China.
Xin CQ; The First People's Hospital of Jingzhou, Jingzhou, Hubei, China.
Fan W; The First Affiliated Hospital of Yangtze University, Jingzhou, Hubei, China.

Front Microbiol ; 14: 1269275, 2023.

Article em En | MEDLINE | ID: mdl-38260899

ABSTRACT

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.