A novel assay based on DNA melting temperature for multiplexed identification of SARS-CoV-2 and influenza A/B viruses

The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and influenza viruses can cause respiratory illnesses with similar clinical symptoms, making their differential diagnoses challenging. Additionally, in critically ill SARS-CoV-2-infected patients, co-infections with other respiratory p...

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Bibliographic Details
Published in:Frontiers in microbiology 2023-12, Vol.14, p.1249085-1249085
Main Authors: Gao, Peng, Fan, Yanyan, Kong, Xiaomu, Zhang, Rui, Chen, Lida, Jiang, Yongwei, Liu, Yi, Zhao, Meimei, Deng, Guoxiong, Cao, Yongtong, Ma, Liang
Format: Article
Language:English
Subjects:
differentiation
influenza viruses
melting curve analysis
Microbiology
SARS-CoV-2
simultaneous detection
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Summary:The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and influenza viruses can cause respiratory illnesses with similar clinical symptoms, making their differential diagnoses challenging. Additionally, in critically ill SARS-CoV-2-infected patients, co-infections with other respiratory pathogens can lead to severe cytokine storm and serious complications. Therefore, a method for simultaneous detection of SARS-CoV-2 and influenza A and B viruses will be clinically beneficial. We designed an assay to detect five gene targets simultaneously via asymmetric PCR-mediated melting curve analysis in a single tube. We used specific probes that hybridize to corresponding single-stranded amplicons at low temperature and dissociate at high temperature, creating different detection peaks representing the targets. The entire reaction was conducted in a closed tube, which minimizes the risk of contamination. The limit of detection, specificity, precision, and accuracy were determined. The assay exhibited a limit of detection of
ISSN:1664-302X
1664-302X
DOI:10.3389/fmicb.2023.1249085