ddPCR: a more accurate tool for SARS-CoV-2 detection in low viral load specimens

• Published in: Emerging microbes & infections Vol. 9; no. 1; pp. 1259 - 1268
• Main Authors: Suo, Tao, Liu, Xinjin, Feng, Jiangpeng, Guo, Ming, Hu, Wenjia, Guo, Dong, Ullah, Hafiz, Yang, Yang, Zhang, Qiuhan, Wang, Xin, Sajid, Muhanmmad, Huang, Zhixiang, Deng, Liping, Chen, Tielong, Liu, Fang, Xu, Ke, Liu, Yuan, Zhang, Qi, Liu, Yingle, Xiong, Yong, Chen, Guozhong, Lan, Ke, Chen, Yu
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 01.01.2020; Taylor & Francis Ltd; Taylor & Francis Group
• Subjects: Betacoronavirus - genetics; clinical detection; Coronavirus Infections - diagnosis; COVID-19; droplet digital PCR; false negative; False Negative Reactions; Humans; Limit of Detection; Pandemics; Pneumonia, Viral - diagnosis; Real-Time Polymerase Chain Reaction - methods; RNA, Viral - genetics; RT-PCR; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2; Viral Load - methods; SARS-CoV-2; false negative; clinical detection; droplet digital PCR; RT-PCR
• ISSN: 2222-1751; 2222-1751
• DOI: 10.1080/22221751.2020.1772678

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, 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 RNA from 77 patients, and compared with RT-PCR in terms of the diagnostic accuracy based on the results of follow-up survey. 26 patients of COVID-19 with negative RT-PCR reports were reported 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 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 RT-PCR.