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. 2020 May 22;1-30.
doi: 10.1080/22221751.2020.1772678. Online ahead of print.

ddPCR: A More Accurate Tool for SARS-CoV-2 Detection in Low Viral Load Specimens

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ddPCR: A More Accurate Tool for SARS-CoV-2 Detection in Low Viral Load Specimens

Tao Suo et al. Emerg Microbes Infect. .
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Abstract

Quantitative real time PCR (RT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. However, due to the low viral load specimens and the limitations of RT-PCR, significant numbers of false negative reports are inevitable, which results in failure to timely diagnose, early treat, cut off transmission, and assess discharge criteria. To improve this situation, an optimized droplet digital PCR (ddPCR) was used for detection of SARS-CoV-2, which showed that the limit of detection of ddPCR is significantly lower than that of RT-PCR. We further explored the feasibility of ddPCR to detect SARS-CoV-2 nucleic acid from 77 clinical throat swabs, and compared with RT-PCR in terms of the diagnostic accuracy based on the results of follow-up survey. According to our study, 26 samples from COVID-19 patients with negative RT-PCR reports were detected as positive by ddPCR. The sensitivity, specificity, PPV, NPV, negative likelihood ratio (NLR) and accuracy were improved from 40% (95% CI: 27-55%), 100% (95% CI: 54-100%), 100%, 16% (95% CI: 13-19%), 0.6 (95% CI: 0.48-0.75) and 47% (95% CI: 33-60%) for RT-PCR to 94% (95% CI: 83-99%), 100% (95% CI: 48-100%), 100%, 63% (95% CI: 36-83%), 0.06 (95% CI: 0.02-0.18) and 95% (95% CI: 84-99%) for ddPCR, respectively. Moreover, 6/14 (42.9%) convalescents were detected as positive by ddPCR, which still carry detectable SARS-CoV-2 at 5-12 days post discharge. Overall, ddPCR shows superiority for clinical diagnosis of SARS-CoV-2 to reduce the false negative reports, which could be a powerful complement to the current standard RT-PCR.

Keywords: RT-PCR; SARS-CoV-2; clinical detection; droplet digital PCR.

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