Novel RT-ddPCR assays for measuring the levels of subgenomic and genomic SARS-CoV-2 transcripts
Autor: | Joseph K. Wong, Ryan Marczak, Albert Vallejo-Gracia, G. Renuka Kumar, Sulggi A. Lee, Holly Anne Martin, Melanie Ott, Steven A. Yukl, Parinaz Fozouni, Sushama Telwatte, Victoria Murray |
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Jazyk: | angličtina |
Rok vydání: | 2021 |
Předmět: |
replication
Viral pathogenesis viruses Messenger Clinical Sciences Computational biology Subgenomic RNA Biology medicine.disease_cause Polymerase Chain Reaction Viral transcription General Biochemistry Genetics and Molecular Biology Article 03 medical and health sciences Transcription (biology) Genetics medicine Humans Digital polymerase chain reaction Viral Guide RNA RNA Messenger skin and connective tissue diseases Lung Molecular Biology 030304 developmental biology Subgenomic mRNA Coronavirus 0303 health sciences SARS-CoV-2 Prevention 030302 biochemistry & molecular biology fungi RNA virus diseases COVID-19 Pneumonia Reverse Transcription Viral transcription/replication Reverse transcriptase body regions Infectious Diseases Droplet digital PCR Quantitative assays RNA Viral Digital PCR Biotechnology |
Zdroj: | Methods (San Diego, Calif.) |
ISSN: | 1095-9130 1046-2023 |
Popis: | 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. |
Databáze: | OpenAIRE |
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