Molecular detection of SARS-CoV-2 using a reagent-free approach

• DOI: 10.1371/journal.pone.0243266
• Publication Date: 2020-12-07T21:51:32Z
• Authors: Ronan Calvez; Andrew Taylor; Leonides Calvo-Bado; Donald Fraser; Colin G. Fink
• Keywords: Consumables; Economic shortage; Sample (material); Molecular diagnostics; Sample Preparation

Shortage of reagents and consumables required for the extraction molecular detection SARS-CoV-2 RNA in respiratory samples has led many laboratories to investigate alternative approaches sample preparation. Many groups recently presented results using heat processing method prior RT-qPCR as an economical enabling extremely fast streamlining processes at virtually no cost. Here, we present our this highlight some major pitfalls that diagnostics should be aware before proceeding with methodology. We first investigated various treatments different temperatures, incubation times volumes optimise treatment conditions. Although initial data confirmed published elsewhere, further investigations revealed unexpected inhibitory properties commonly used universal transport media (UTMs) on commercially available mixes, leading a risk reporting false-negative results. This emphasises critical importance thorough validation process determine most suitable use depending types tested. In conclusion, is effective very consistent Ct values sensitivity 96.2% when compared conventional method. It also include internal control check each potential inhibition.