4.7 Article

A novel One-pot rapid diagnostic technology for COVID-19

Journal

ANALYTICA CHIMICA ACTA
Volume 1154, Issue -, Pages -

Publisher

ELSEVIER
DOI: 10.1016/j.aca.2021.338310

Keywords

COVID-19; RT-qPCR; RT-LAMP; One-pot reaction

Funding

  1. Guangdong Science and Technology Project [2020A111128016]
  2. Special project for emergency research of Dongguan City [2020717102119]
  3. Guangdong Medical Research Fund Project [A2020006]

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A novel one-tube rapid diagnostic approach based on formamide and colorimetric RT-LAMP was established for the detection of SARS-CoV-2 without the extraction of specific viral RNA. The technique showed high specificity and could visually detect the virus within 45 minutes.
Novel coronavirus disease (COVID-19) caused by SARS-CoV-2 is an ongoing global pandemic associated with high rates of morbidity and mortality. RT-qPCR has become the diagnostic standard for the testing of SARS-CoV-2 in most countries. COVID-19 diagnosis generally relies upon RT-qPCR-mediated identification of SARS-CoV-2 viral RNA, which is costly, labor-extensive, and requires specialized training and equipment. Herein, we established a novel one-tube rapid diagnostic approach based upon formamide and colorimetric RT-LAMP (One-Pot RT-LAMP) that can be used to diagnose COVID-19 without the extraction of specific viral RNA. The technique could visually detect SARS-CoV-2 within 45 min with a limit of detection of 5 copies per reaction in extracted RNA, and about 7.66 virus copies per mu L in viral transport medium. The One-Pot RT-LAMP test showed a high specificity without cross-reactivity with 12 viruses including SARS-CoV, MERS-CoV, and human infectious influenza virus (H1N1/H3N2 of influenza A and B virus, ect. We validated this One-Pot RT-LAMP approach by its successful use for the analysis of 45 clinical nasopharyngeal swab samples, yielding results identical to those of traditional RT-qPCR analyses, while achieving good selectivity and sensitivity relative to a commercial RT-qPCR approach. As such, this One-Pot RT-LAMP technology may be a valid means of conducting high-sensitivity, low-cost and rapid SARS-CoV-2 identification without the extraction of viral RNA. (C) 2021 Elsevier B.V. All rights reserved.

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