Rapid Detection of Novel Coronavirus (COVID-19) by Reverse Transcription-Loop-Mediated Isothermal Amplification

Laura E Lamb; Sarah N Bartolone; Elijah Ward; Michael B Chancellor

This study has several limitations. First, COVID-19 is Biosafety level 3 so our laboratory was unable to work directly with the virus or with infected samples. As such, all the experiments presented here used a nucleotide oligo of COVID-19 corresponding to the GenBank MN908947 sequence. Similarly, we were unable to directly test related coronaviruses and instead used nucleotide oligos from the same region of those viruses. However, these do represent a proof-of-feasibility for this assay, and the primers were further evaluated for specificity by BLASTing them to related coronavirus sequences. Although RT-LAMP reactions are highly specific, it is not a quantitative test. However other groups are working at improving the read outs of RT-LAMP assays including the use of smartphone-integrated sensors to make interpretation of the assay even more user-friendly. 8 RT-LAMP reactions can have a higher rate of false positives compared to qRT-PCR; we did not experience this in any of our no template negative control reactions, negative control samples, or other samples containing other coronaviruses. We also took the All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity.