Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-qPCR

Ioanna Smyrlaki; Martin Ekman; Martin Vondracek; Natali Papanicoloau; Antonio Lentini; Johan Aarum; Shaman Muradrasoli; Jan Albert; Björn Högberg; Björn Reinius

Scaled, rapid and affordable COVID-19 diagnostics could help to limit the spread of SARS-CoV-2, potentially saving many of lives, but RNA extraction constitutes a barrier to scale-up of testing. We explored methods to circumvent RNA extraction by performing RT-qPCR directly on heat-inactivated subject samples and sample lysates. Our results show that RT-qPCR-based testing for SARS-CoV-2 infection can be performed through significantly simpler protocols and without the use of RNA extraction kits, nor other special kits. The results also suggest that this might be achieved without any major sacrifice in accuracy of determining negative and positive cases. The procedure could be especially useful for massively scaling up SARS-CoV-2 testing, as the logistics and cost of RNA purification could be unworkable in mass testing. Importantly, the direct method is also attractive in settings where repeated, cheaper, and All rights reserved. No reuse allowed without permission. the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.