tetano
Editor, Senior Moderator
Anal Chim Acta
. 2021 Apr 15;1154:338310.
doi: 10.1016/j.aca.2021.338310. Epub 2021 Feb 11.
A novel One-pot rapid diagnostic technology for COVID-19
Junmin Li[SUP] 1 [/SUP], Xuejiao Hu[SUP] 2 [/SUP], Xiaoming Wang[SUP] 1 [/SUP], Jianing Yang[SUP] 1 [/SUP], Lei Zhang[SUP] 1 [/SUP], Qianyun Deng[SUP] 2 [/SUP], Xiqin Zhang[SUP] 2 [/SUP], Zixia Wang[SUP] 2 [/SUP], Tieying Hou[SUP] 3 [/SUP], Shan Li[SUP] 4 [/SUP]
Affiliations
Abstract
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 ?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.
Keywords: COVID-19; One-pot reaction; RT-LAMP; RT-qPCR.
. 2021 Apr 15;1154:338310.
doi: 10.1016/j.aca.2021.338310. Epub 2021 Feb 11.
A novel One-pot rapid diagnostic technology for COVID-19
Junmin Li[SUP] 1 [/SUP], Xuejiao Hu[SUP] 2 [/SUP], Xiaoming Wang[SUP] 1 [/SUP], Jianing Yang[SUP] 1 [/SUP], Lei Zhang[SUP] 1 [/SUP], Qianyun Deng[SUP] 2 [/SUP], Xiqin Zhang[SUP] 2 [/SUP], Zixia Wang[SUP] 2 [/SUP], Tieying Hou[SUP] 3 [/SUP], Shan Li[SUP] 4 [/SUP]
Affiliations
- PMID: 33736798
- DOI: 10.1016/j.aca.2021.338310
Abstract
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 ?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.
Keywords: COVID-19; One-pot reaction; RT-LAMP; RT-qPCR.