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. 2020 Dec:286:113965.
doi: 10.1016/j.jviromet.2020.113965. Epub 2020 Sep 3.

Rapid, convenient and efficient kit-independent detection of SARS-CoV-2 RNA

Detlef Michel  1 , Karin M Danzer  2 , Rüdiger Groß  3 , Carina Conzelmann  3 , Janis A Müller  3 , Axel Freischmidt  4 , Jochen H Weishaupt  4 , Sandra Heller  5 , Jan Münch  3 , Manuela Michel  1 , Thomas Stamminger  1 , Alexander Kleger  6 , Markus Otto  4
Affiliations

Rapid, convenient and efficient kit-independent detection of SARS-CoV-2 RNA

Detlef Michel et al. J Virol Methods. 2020 Dec.

Abstract

Pandemic SARS-CoV-2 infection has rapidly developed into a socioeconomic and humanitarian catastrophe. Basic principles to prevent SARS-CoV-2 transmission are social distancing, face masks, contact tracing and early detection of SARS-CoV-2. To meet these requirements, virtually unlimited test capacities delivering results in a rapid and reliable manner are a prerequisite. Here, we provide and validate such a rapid, convenient and efficient kit-independent detection of SARS-CoV-2 RNA, termed COVID-quick-DET. This straightforward method operates with simple proteinase K treatment and repetitive heating steps with a sensitivity of 94.6% in head-to-head comparisons with kit-based isolation methods. This result is supported by data obtained from serially diluted SARS-CoV-2 virus stocks. Given its cost- and time-effective operation, COVID-quick-DET might be best suited for countries with general shortage or temporary acute scarcity of resources and equipment.

Keywords: COVID-19; Heating; Kit-free RNA extraction; Proteinase K; SARS-CoV-2.

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Conflict of interest statement

The authors do not have any conflict of interest

Figures

Fig. 1
Fig. 1
(A) Heat map of SARS-CoV-2 gene equivalents (log10) for 56 COVID-19 patients using a commercial RNA isolation kit or COVID-quick-DET. The colors in the heat map represent gene expression levels (log10). White crossed boxes marked with “#” represent the samples giving discrepant results between COVID-quick-DET and the reference method. Samples taken from bronchoalveolar lavages are indicated by asterisk. (B) Time for RNA isolation per 100 samples in minutes. (C) Costs for RNA isolation per 100 samples in Euro. (D) Schematic workflow of tested material compatible with COVID-Quick-DET method and successful qPCR. (E) Detection of SARS-CoV-2 ORF1b-nsp14 in samples prepared by serial dilution of virus stocks of two strains in medium (French-left panel; Dutch-right panel). Viral RNA was either extracted by Qiagen Viral RNA Mini Kit (Qiagen #52906) or prepared by the COVID-quick-DET protocol before being subjected to RT-qPCR as described (Chu et al., 2020; https://www.who.int/). For direct comparison to COVID-quick-DET, RNA was eluted in the original sample volume without concentration using Qiagen Viral RNA Mini Kit. Ct 35 was set as cut-off. Reactions have been run in duplicates, values are mean ± SD. N.d. = not detected (Ct => 35).
See this image and copyright information in PMC

References

    1. Bai Y., Xiao Y., Suo Y., Shen Y., Shao Y., Zhang D., Zhou C. Enhancement of PCR sensitivity and yield using thiol-modified primers. Sci. Rep. 2018;8:14858. - PMC - PubMed
    1. Chu D.K.W., Pan Y., Cheng S.M.S., Hui K.P.Y., Krishnan P., Liu Y., Ng D.Y.M., Wan C.K.C., Yang P., Wang Q., Peiris M., Poon L.L.M. Molecular diagnosis of a novel coronavirus (2019-nCoV) causing an outbreak of pneumonia. Clin. Chem. 2020;66:549–555. - PMC - PubMed
    1. Corman V.M., Landt O., Kaiser M., Molenkamp R., Meijer A., Chu D.K., Bleicker T., Brünink S., Schneider J., Schmidt M.L., Mulders D.G., Haagmans B.L., van der Veer B., van den Brink S., Wijsman L., Goderski G., Romette J.-L., Ellis J., Zambon M., Peiris M., Goossens H., Reusken C., Koopmans M.P., Drosten C. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Eurosurveillance. 2020;25 2000045. - PMC - PubMed
    1. Darnell M.E., Subbarao K., Feinstone S.M., Taylor D.R. Inactivation of the coronavirus that induces severe acute respiratory syndrome, SARS-CoV. J. Virol. Methods. 2004;121:85–91. - PMC - PubMed
    1. Fiore V.G., DeFelice N., Glicksberg B.S., Perl O., Shuster A., Kulkarni K., O’Brien M., Pisauro M.A., Chung D., Gu X. 2020. Containment of Future Waves of COVID-19: Simulating the Impact of Different Policies and Testing Capacities for Contact Tracing, Testing, and Isolation. medRxiv. - PMC - PubMed

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