Digital Microfluidic Multiplex RT-qPCR for SARS-CoV-2 Detection and Variants Discrimination

Continuous mutations have occurred in the genome of the SARS-CoV-2 virus since the onset of the COVID-19 pandemic. The increased transmissibility of the mutated viruses has not only imposed medical burdens but also prolonged the duration of the pandemic. A point-of-care (POC) platform that provides...

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Bibliographic Details
Published in:Micromachines (Basel) 2023-08, Vol.14 (8), p.1627
Main Authors: Ho, Kuan-Lun, Ding, Jing, Fan, Jia-Shao, Tsui, Wai Ning Tiffany, Bai, Jianfa, Fan, Shih-Kang
Format: Article
Language:English
Subjects:
Antigens
COVID-19
COVID-19 vaccines
Delta variant
digital microfluidics
Electrodes
Genes
Genomes
Genomics
Health aspects
Infections
Medical research
Microfluidics
Omicron variant
Pandemics
Polymerase chain reaction
RNA
RT-qPCR
SARS-CoV-2
Serology
Severe acute respiratory syndrome coronavirus 2
Viral diseases
Viral infections
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Summary:Continuous mutations have occurred in the genome of the SARS-CoV-2 virus since the onset of the COVID-19 pandemic. The increased transmissibility of the mutated viruses has not only imposed medical burdens but also prolonged the duration of the pandemic. A point-of-care (POC) platform that provides multitarget detection will help to track and reduce disease transmissions. Here we detected and discriminated three genotypes of SARS-CoV-2, including the wildtype and two variants of concern (VOCs), the Delta variant and Omicron variant, through reverse transcription quantitative polymerase chain reaction (RT-qPCR) on a digital microfluidics (DMF)-based cartridge. Upon evaluating with the RNA samples of Omicron variant, the DMF RT-qPCR presented a sensitivity of 10 copies/μL and an amplification efficiency of 96.1%, capable for clinical diagnosis. When spiking with SARS-CoV-2 RNA (wildtype, Delta variant, or Omicron variant) and 18S rDNA, the clinical analog samples demonstrated accurate detection and discrimination of different SARS-CoV-2 strains in 49 min.
ISSN:2072-666X
2072-666X
DOI:10.3390/mi14081627