Development of multiplex RT-ddPCR assays for detection of SARS-CoV-2 and other common respiratory virus infections.

Autor: Leong NKC; School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong., Gu H; School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong., Ng DYM; School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong., Chang LDJ; School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong., Krishnan P; School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong., Cheng SSM; School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong., Peiris M; School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong.; Centre for Immunology and Infection, Hong Kong Science and Technology Park, Hong Kong.; HKU-Pasteur Research Pole, School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong., Poon LLM; School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong.; Centre for Immunology and Infection, Hong Kong Science and Technology Park, Hong Kong.; HKU-Pasteur Research Pole, School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong.
Jazyk: angličtina
Zdroj: Influenza and other respiratory viruses [Influenza Other Respir Viruses] 2023 Jan; Vol. 17 (1), pp. e13084. Date of Electronic Publication: 2022 Dec 14.
DOI: 10.1111/irv.13084
Abstrakt: Background: Measures for mitigation of Coronavirus Disease 2019 (COVID-19) were set to reduce the spread of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). SARS-CoV-2 and other respiratory viruses share similar transmission routes and some common clinical manifestations. Co-circulation of SARS-CoV-2 and other common respiratory viruses is imminent. Therefore, development of multiplex assays for detecting these respiratory viruses is essential for being prepared for future outbreaks of respiratory viruses.
Methods: A panel of three reverse transcription droplet digital PCR (RT-ddPCR) assays were developed to detect 15 different human respiratory viruses. Evaluations of its performance were demonstrated. A total of 100 local and 98 imported COVID-19 cases in Hong Kong were screened for co-infection with other common respiratory viruses.
Results: All detected viral targets showed distinct signal clusters using the multiplex RT-ddPCR assays. These assays have a broad range of linearity and good intra-/inter-assay reproducibility for each target. The lower limits of quantification for all targets were ≤46 copies per reaction. Six imported cases of COVID-19 were found to be co-infected with other respiratory viruses, whereas no local case of co-infection was observed.
Conclusions: The multiplex RT-ddPCR assays were demonstrated to be useful for screening of respiratory virus co-infections. The strict preventive measures applied in Hong Kong may be effective in limiting the circulation of other human respiratory viruses. The multiplex assays developed in this study can achieve a robust detection method for clinical and research purposes.
(© 2022 The Authors. Influenza and Other Respiratory Viruses published by John Wiley & Sons Ltd.)
Databáze: MEDLINE
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