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

• 出版年: Micromachines
• 主要な著者: Kuan-Lun Ho, Jing Ding, Jia-Shao Fan, Wai Ning Tiffany Tsui, Jianfa Bai, Shih-Kang Fan
• フォーマット: 論文
• 言語: 英語
• 出版事項: MDPI AG 2023-08-01
• 主題: SARS-CoV-2; COVID-19; Delta variant; Omicron variant; RT-qPCR; digital microfluidics
• オンライン･アクセス: https://www.mdpi.com/2072-666X/14/8/1627

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.