Open AccessSuitcase Lab for Rapid Detection of SARS-CoV-2 Based on Recombinase Polymerase Amplification Assay
Author(s) -
Ahmed Abd El Wahed,
Pranav Patel,
Melanie Maier,
Corinna Pietsch,
Dana Rüster,
Susanne Böhlken-Fascher,
Jonas Kissenkötter,
Ole Behrmann,
Michael Frimpong,
Moussa Moïse Diagne,
Martin Faye,
Ndongo Dia,
Mohamed A. Shalaby,
Haitham M. Amer,
Mahmoud A. ElGamal,
Aya Zaki,
Ghada Ismail,
Marco Kaiser,
Victor M. Corman,
Matthias Niedrig,
Olfert Landt,
Ousmane Faye,
Amadou Alpha Sall,
Frank T. Hufert,
Uwe Truyen,
Uwe G. Liebert,
Manfred Weidmann
Publication year - 2021
Publication title -
analytical chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.117
H-Index - 332
eISSN - 1520-6882
pISSN - 0003-2700
DOI - 10.1021/acs.analchem.0c04779
Subject(s) - recombinase polymerase amplification , virology , microbiology and biotechnology , chemistry , rna , nucleic acid , rna polymerase , polymerase , rna dependent rna polymerase , polymerase chain reaction , recombinase , real time polymerase chain reaction , gene , biology , biochemistry , recombination
In March 2020, the SARS-CoV-2 virus outbreak was declared as a world pandemic by the World Health Organization (WHO). The only measures for controlling the outbreak are testing and isolation of infected cases. Molecular real-time polymerase chain reaction (PCR) assays are very sensitive but require highly equipped laboratories and well-trained personnel. In this study, a rapid point-of-need detection method was developed to detect the RNA-dependent RNA polymerase (RdRP), envelope protein (E), and nucleocapsid protein (N) genes of SARS-CoV-2 based on the reverse transcription recombinase polymerase amplification (RT-RPA) assay. RdRP, E, and N RT-RPA assays required approximately 15 min to amplify 2, 15, and 15 RNA molecules of molecular standard/reaction, respectively. RdRP and E RT-RPA assays detected SARS-CoV-1 and 2 genomic RNA, whereas the N RT-RPA assay identified only SARS-CoV-2 RNA. All established assays did not cross-react with nucleic acids of other respiratory pathogens. The RT-RPA assay's clinical sensitivity and specificity in comparison to real-time RT-PCR ( n = 36) were 94 and 100% for RdRP; 65 and 77% for E; and 83 and 94% for the N RT-RPA assay. The assays were deployed to the field, where the RdRP RT-RPA assays confirmed to produce the most accurate results in three different laboratories in Africa ( n = 89). The RPA assays were run in a mobile suitcase laboratory to facilitate the deployment at point of need. The assays can contribute to speed up the control measures as well as assist in the detection of COVID-19 cases in low-resource settings.