Department of Biosciences, Biotechnology and Biopharmaceutics, University of Bari Aldo Moro, Via Orabona 4, 70126, Bari, Italy.
Institute of Biomembranes, Bioenergetics and Molecular Biotechnologies, National Research Council, Via Amendola 122/O, 70126, Bari, Italy.
Commun Biol. 2021 Oct 22;4(1):1215. doi: 10.1038/s42003-021-02748-0.
SARS-CoV-2 replication requires the synthesis of a set of structural proteins expressed through discontinuous transcription of ten subgenomic mRNAs (sgmRNAs). Here, we have fine-tuned droplet digital PCR (ddPCR) assays to accurately detect and quantify SARS-CoV-2 genomic ORF1ab and sgmRNAs for the nucleocapsid (N) and spike (S) proteins. We analyzed 166 RNA samples from anonymized SARS-CoV-2 positive subjects and we observed a recurrent and characteristic pattern of sgmRNAs expression in relation to the total viral RNA content. Additionally, expression profiles of sgmRNAs, as determined by meta-transcriptomics sequencing of a subset of 110 RNA samples, were highly correlated with those obtained by ddPCR. By providing a comprehensive and dynamic snapshot of the levels of SARS-CoV-2 sgmRNAs in infected individuals, our results may contribute a better understanding of the dynamics of transcription and expression of the genome of SARS-CoV-2 and facilitate the development of more accurate molecular diagnostic tools for the stratification of COVID-19 patients.
SARS-CoV-2 的复制需要通过十个亚基因组 mRNA(sgmRNAs)的不连续转录来合成一组结构蛋白。在这里,我们对液滴数字 PCR(ddPCR)检测进行了微调,以准确检测和定量 SARS-CoV-2 基因组 ORF1ab 和核衣壳(N)和刺突(S)蛋白的 sgmRNAs。我们分析了来自匿名 SARS-CoV-2 阳性受试者的 166 个 RNA 样本,观察到 sgmRNAs 的表达与总病毒 RNA 含量之间存在反复出现的特征性模式。此外,通过对 110 个 RNA 样本的亚转录组学测序确定的 sgmRNAs 表达谱与 ddPCR 获得的表达谱高度相关。通过提供受感染个体中 SARS-CoV-2 sgmRNAs 水平的全面和动态快照,我们的结果可能有助于更好地理解 SARS-CoV-2 基因组转录和表达的动态,并促进开发更准确的分子诊断工具,以对 COVID-19 患者进行分层。