A DNA intercalating dye-based RT-qPCR alternative to diagnose SARS-CoV-2

FEDERICO FUCHS WIGHTMAN; JOSÉ N. STIGLIANO; MARCOS PALAVECINO; JOSÉ CLEMENTE; ESTEFANÍA BENEDETTI; FEREICO REMES LENICOV; ALBERTO R. KORNBLIHTT; MANUEL JAVIER MUÑOZ; MICAELA A. GODOY HERZ; LAUREANO BRAGADO; LUCAS SERVI; MARTÍN AVARO; ELSA BAUMEISTER; CYBELE GARCIA ; ANABELLA SREBROW; IGNACIO E. SCHOR; MUÑOZ, JUAN CRISTÓBAL; NICOLAS NIETO MORENO; GONZALO CABRERIZO; ANDREA PONTORIERO; FABIAN RUDOLF; VALERIA BUGGIANO; MANUEL DE LA MATA; EZEQUIEL PETRILLO

RNA BIOLOGY

LANDES BIOSCIENCE

Lugar: Austin, Texas; Año: 2021 p. 2218 - 2225

1547-6286

Early detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been proven crucial during the efforts to mitigate the effects of the COVID-19 pandemic. Several diagnostic methods have emerged in the past few months, each with different shortcomings and limitations. The current gold standard, RT-qPCR using fluorescent probes, relies on demanding equipment requirements plus the high costs of the probes and specific reaction mixes. To broaden the possibilities of reagents and thermocyclers that could be allocated towards this task, we have optimized an alternative strategy for RT-qPCR diagnosis. This is based on a widely used DNA-intercalating dye and can be implemented with several different qPCR reagents and instruments. Remarkably, the proposed qPCR method performs similarly to the broadly used TaqMan-based detection, in terms of specificity and sensitivity, thus representing a reliable tool. We think that, through enabling the use of vast range of thermocycler models and laboratory facilities for SARS-CoV-2 diagnosis, the alternative proposed here can increase dramatically the testing capability, especially in countries with limited access to costly technology and reagents.