tetano
Editor, Senior Moderator
Talanta
. 2021 Mar 1;224:121726.
doi: 10.1016/j.talanta.2020.121726. Epub 2020 Oct 27.
Highly accurate and sensitive diagnostic detection of SARS-CoV-2 by digital PCR
Lianhua Dong[SUP] 1 [/SUP], Junbo Zhou[SUP] 2 [/SUP], Chunyan Niu[SUP] 1 [/SUP], Quanyi Wang[SUP] 3 [/SUP], Yang Pan[SUP] 3 [/SUP], Sitong Sheng[SUP] 4 [/SUP], Xia Wang[SUP] 1 [/SUP], Yongzhuo Zhang[SUP] 1 [/SUP], Jiayi Yang[SUP] 1 [/SUP], Manqing Liu[SUP] 2 [/SUP], Yang Zhao[SUP] 1 [/SUP], Xiaoying Zhang[SUP] 4 [/SUP], Tao Zhu[SUP] 5 [/SUP], Tao Peng[SUP] 1 [/SUP], Jie Xie[SUP] 1 [/SUP], Yunhua Gao[SUP] 1 [/SUP], Di Wang[SUP] 1 [/SUP], Xinhua Dai[SUP] 6 [/SUP], Xiang Fang[SUP] 7 [/SUP]
Affiliations
Abstract
The outbreak of COVID-19 caused by a novel Coronavirus (termed SARS-CoV-2) has spread to over 210 countries around the world. Currently, reverse transcription quantitative qPCR (RT-qPCR) is used as the gold standard for diagnosis of SARS-CoV-2. However, the sensitivity of RT-qPCR assays of pharyngeal swab samples are reported to vary from 30% to 60%. More accurate and sensitive methods are urgently needed to support the quality assurance of the RT-qPCR or as an alternative diagnostic approach. A reverse transcription digital PCR (RT-dPCR) method was established and evaluated. To explore the feasibility of RT-dPCR in diagnostic of SARS-CoV-2, a total of 196 clinical pharyngeal swab samples from 103 suspected patients, 77 close contacts and 16 supposed convalescents were analyzed by RT-qPCR and then measured by the proposed RT-dPCR. For the 103 fever suspected patients, 19 (19/25) negative and 42 (42/49) equivocal tested by RT-qPCR were positive according to RT-dPCR. The sensitivity of SARS-CoV-2 detection was significantly improved from 28.2% by RT-qPCR to 87.4% by RT-dPCR. For 29 close contacts (confirmed by additional sample and clinical follow up), 16 (16/17) equivocal and 1 negative tested by RT-qPCR were positive according to RT-dPCR, which is implying that the RT-qPCR is missing a lot of asymptomatic patients. The overall sensitivity, specificity and diagnostic accuracy of RT-dPCR were 91%, 100% and 93%, respectively. RT-dPCR is highly accurate method and suitable for detection of pharyngeal swab samples from COVID-19 suspected patients and patients under isolation and observation who may not be exhibiting clinical symptoms.
Keywords: Coronavirus disease (COVID-19); Novel coronavirus (termed SARS-CoV-2); Reverse transcription digital PCR (RT-dPCR); Reverse transcription quantitative qPCR (RT-qPCR); Sensitivity.
. 2021 Mar 1;224:121726.
doi: 10.1016/j.talanta.2020.121726. Epub 2020 Oct 27.
Highly accurate and sensitive diagnostic detection of SARS-CoV-2 by digital PCR
Lianhua Dong[SUP] 1 [/SUP], Junbo Zhou[SUP] 2 [/SUP], Chunyan Niu[SUP] 1 [/SUP], Quanyi Wang[SUP] 3 [/SUP], Yang Pan[SUP] 3 [/SUP], Sitong Sheng[SUP] 4 [/SUP], Xia Wang[SUP] 1 [/SUP], Yongzhuo Zhang[SUP] 1 [/SUP], Jiayi Yang[SUP] 1 [/SUP], Manqing Liu[SUP] 2 [/SUP], Yang Zhao[SUP] 1 [/SUP], Xiaoying Zhang[SUP] 4 [/SUP], Tao Zhu[SUP] 5 [/SUP], Tao Peng[SUP] 1 [/SUP], Jie Xie[SUP] 1 [/SUP], Yunhua Gao[SUP] 1 [/SUP], Di Wang[SUP] 1 [/SUP], Xinhua Dai[SUP] 6 [/SUP], Xiang Fang[SUP] 7 [/SUP]
Affiliations
- PMID: 33379001
- PMCID: PMC7588801
- DOI: 10.1016/j.talanta.2020.121726
Abstract
The outbreak of COVID-19 caused by a novel Coronavirus (termed SARS-CoV-2) has spread to over 210 countries around the world. Currently, reverse transcription quantitative qPCR (RT-qPCR) is used as the gold standard for diagnosis of SARS-CoV-2. However, the sensitivity of RT-qPCR assays of pharyngeal swab samples are reported to vary from 30% to 60%. More accurate and sensitive methods are urgently needed to support the quality assurance of the RT-qPCR or as an alternative diagnostic approach. A reverse transcription digital PCR (RT-dPCR) method was established and evaluated. To explore the feasibility of RT-dPCR in diagnostic of SARS-CoV-2, a total of 196 clinical pharyngeal swab samples from 103 suspected patients, 77 close contacts and 16 supposed convalescents were analyzed by RT-qPCR and then measured by the proposed RT-dPCR. For the 103 fever suspected patients, 19 (19/25) negative and 42 (42/49) equivocal tested by RT-qPCR were positive according to RT-dPCR. The sensitivity of SARS-CoV-2 detection was significantly improved from 28.2% by RT-qPCR to 87.4% by RT-dPCR. For 29 close contacts (confirmed by additional sample and clinical follow up), 16 (16/17) equivocal and 1 negative tested by RT-qPCR were positive according to RT-dPCR, which is implying that the RT-qPCR is missing a lot of asymptomatic patients. The overall sensitivity, specificity and diagnostic accuracy of RT-dPCR were 91%, 100% and 93%, respectively. RT-dPCR is highly accurate method and suitable for detection of pharyngeal swab samples from COVID-19 suspected patients and patients under isolation and observation who may not be exhibiting clinical symptoms.
Keywords: Coronavirus disease (COVID-19); Novel coronavirus (termed SARS-CoV-2); Reverse transcription digital PCR (RT-dPCR); Reverse transcription quantitative qPCR (RT-qPCR); Sensitivity.