Journal article
Novel RT-ddPCR assays for simultaneous quantification of multiple noncoding and coding regions of SARS-CoV-2 RNA
S Telwatte, N Kumar, A Vallejo-Gracia, GR Kumar, CM Lu, M Ott, JK Wong, SA Yukl
Journal of Virological Methods | Published : 2021
Abstract
A hallmark of coronavirus transcription is the generation of negative-sense RNA intermediates that serve as the templates for the synthesis of positive-sense genomic RNA (gRNA) and an array of subgenomic mRNAs (sgRNAs) encompassing sequences arising from discontinuous transcription. Existing PCR-based diagnostic assays for SAR-CoV-2 are qualitative or semi-quantitative and do not provide the resolution needed to assess the complex transcription dynamics of SARS-CoV-2 over the course of infection. We developed and validated a novel panel of sensitive, quantitative RT-ddPCR assays designed to target regions spanning the genome of SARS-CoV-2. Our assays target untranslated regions (5′, 3′) as w..
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Awarded by National Institutes of Health
Funding Acknowledgements
This research was supported by funds from the Emergency COVID-19 Research Seed Funding of the University of California (Grant Number R00RG3113 [ST]). The investigators received salary support from the U.S. Department of Veterans Affairs (SAY and JKW), the National Institute of Diabetes and Digestive and Kidney Diseases at the NIH (R01DK108349 [SAY, JKW], R01DK120387 [SAY]), the National Institute of Allergy and Infectious Diseases at the NIH (R01AI132128 [SAY, JKW]), the UCSF/GIVI Center for AIDS Research (CFAR; Grant#P30 AI027763 [ST]; award #A120163 [PI: Paul Volberding]), and the California HIV/AIDS Research Program (Grant number BB19-SF-009 [ST]). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.