Abstract
The replication of SARS-CoV-2 and other coronaviruses depends on transcription of negative-sense RNA intermediates that serve as the templates for the synthesis of positive-sense genomic RNA (gRNA) and multiple different subgenomic mRNAs (sgRNAs) encompassing fragments arising from discontinuous transcription. Recent studies have aimed to characterize the expression of subgenomic SARS-CoV-2 transcripts in order to investigate their clinical significance. Here, we describe a novel panel of reverse transcription droplet digital PCR (RT-ddPCR) assays designed to specifically quantify multiple different subgenomic SARS-CoV-2 transcripts and distinguish them from transcripts that do not arise from discontinuous transcription at each locus. These assays can be applied to samples from SARS-CoV-2 infected patients to better understand the regulation of SARS-CoV-2 transcription and how different sgRNAs may contribute to viral pathogenesis and clinical disease severity.
Keywords: COVID-19; Coronavirus; Digital PCR; Droplet digital PCR; Quantitative assays; SARS-CoV-2; Subgenomic RNA; Viral transcription/replication.
【저자키워드】 COVID-19, SARS-CoV-2, coronavirus, digital PCR, droplet digital PCR, subgenomic RNA, Quantitative assays, Viral transcription/replication., 【초록키워드】 severity, Transcription, viral pathogenesis, droplet, RNA, Replication, digital PCR, Digital, Clinical significance, sgRNA, reverse transcription, synthesis, discontinuous transcription, expression, Quantitative assays, subgenomic mRNA, genomic RNA, Regulation, locus, SARS-CoV-2 infected patients, subgenomic mRNAs, other coronaviruses, transcripts, sgRNAs, clinical disease, gRNA, viral transcription, replication of SARS-CoV-2, transcript, recent, positive-sense, arising, other coronavirus, contribute, SARS-CoV-2 infected patient, 【제목키워드】 novel, genomic, transcript,