Tracking the Transcription Kinetic of SARS-CoV-2 in Human Cells by Reverse Transcription-Droplet Digital PCR.

Autor: Au KK; Medtimes Molecular Laboratory Limited, Hong Kong, China., Chen C; The Jockey Club School of Public Health and Primary Care, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China.; Li Ka Shing Institute of Health Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China., Chan YM; Medtimes Molecular Laboratory Limited, Hong Kong, China., Wong WWS; Medtimes Molecular Laboratory Limited, Hong Kong, China., Lv H; HKU-Pasteur Research Pole, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China., Mok CKP; The Jockey Club School of Public Health and Primary Care, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China.; Li Ka Shing Institute of Health Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China., Chow CK; Medtimes Molecular Laboratory Limited, Hong Kong, China.
Jazyk: angličtina
Zdroj: Pathogens (Basel, Switzerland) [Pathogens] 2021 Oct 02; Vol. 10 (10). Date of Electronic Publication: 2021 Oct 02.
DOI: 10.3390/pathogens10101274
Abstrakt: Viral transcription is an essential step of SARS-CoV-2 infection after invasion into the target cells. Antiviral drugs such as remdesivir, which is used to treat COVID-19 patients, targets the viral RNA synthesis. Understanding the mechanism of viral transcription may help to develop new therapeutic treatment by perturbing virus replication. In this study, we established 28 ddPCR assays and designed specific primers/probe sets to detect the RNA levels of 15 NSP, 9 ORF, and 4 structural genes of SARS-CoV-2. The transcriptional kinetics of these viral genes were determined longitudinally from the beginning of infection to 12 h postinfection in Caco-2 cells. We found that SARS-CoV-2 takes around 6 h to hijack the cells before the initiation of viral transcription process in human cells. Our results may contribute to a deeper understanding of the mechanisms of SARS-CoV-2 infection.
Databáze: MEDLINE