A Comparative Study of Real-Time RT-PCR-Based SARS-CoV-2 Detection Methods and Its Application to Human-Derived and Surface Swabbed Material.
A Comparative Study of Real-Time RT-PCR-Based SARS-CoV-2 Detection Methods and Its Application to Human-Derived and Surface Swabbed Material.
复制标题
DOI：
10.1016/j.jmoldx.2021.04.009
复制
发表时间：
2021-07
期刊：
影响因子：
--
通讯作者：
Levesque MP
中科院分区：
文献类型：
--
作者：
Tastanova A;Stoffel CI;Dzung A;Cheng PF;Bellini E;Johansen P;Duda A;Nobbe S;Lienhard R;Bosshard PP;Levesque MP
实时RT-PCR仍然是检测各种病毒性疾病的金标准。在2019年冠状病毒大流行中，开发了多种基于RT-PCR的测试来筛查病毒感染。作为对日益增长的检测需求的应急响应，我们建立了一个严重急性呼吸综合征冠状病毒2（SARS-CoV-2）PCR诊断平台，并比较了不同的商业和内部RT-PCR方案。用92份SARS-CoV-2阳性和92份SARS-CoV-2阴性样本评价了4种商业RT-PCR方案、1种定制RT-PCR方案和1种内部RT-PCR方案。并对这些方案的经济性和实用性进行了比较。此外，还建立了一种高灵敏度的数字液滴PCR（ddPCR）方法，并对RT-PCR和ddPCR方法在SARS-CoV-2环境样品中的应用进行了研究。结果表明，两种RT-PCR方法的检测限均较低（1 ~ 2个病毒拷贝/μL），灵敏度和特异性分别为93.6%~ 97.8%和98.7%~ 100%，并证明了两种商业化方法的可行性。测试的商业和定制的RT-PCR检测试剂盒显示出非常好的和可比的灵敏度和特异性，试剂盒可以进一步优化用于来自人类和表面拭子样本的SARS-CoV-2病毒样本。
Real-time RT-PCR remains a gold standard in the detection of various viral diseases. In the coronavirus 2019 pandemic, multiple RT-PCR–based tests were developed to screen for viral infection. As an emergency response to increasing testing demand, we established a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) PCR diagnostics platform for which we compared different commercial and in-house RT-PCR protocols. Four commercial, one customized, and one in-house RT-PCR protocols were evaluated with 92 SARS-CoV-2–positive and 92 SARS-CoV-2–negative samples. Furthermore, economical and practical characteristics of these protocols were compared. In addition, a highly sensitive digital droplet PCR (ddPCR) method was developed, and application of RT-PCR and ddPCR methods on SARS-CoV-2 environmental samples was examined. Very low limits of detection (1 or 2 viral copies/μL), high sensitivities (93.6% to 97.8%), and high specificities (98.7% to 100%) for the tested RT-PCR protocols were found. Furthermore, the feasibility of downscaling two of the commercial protocols, which could optimize testing capacity, was demonstrated. Tested commercial and customized RT-PCR detection kits show very good and comparable sensitivity and specificity, and the kits could be further optimized for use on SARS-CoV-2 viral samples derived from human and surface swabbed samples.
登录
查看更多内容
DOI：
10.1016/j.jcv.2020.104510
发表时间：
2020-08
期刊：
Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology
影响因子：
--
作者：
Iglói Z;Leven M;Abdel-Karem Abou-Nouar Z;Weller B;Matheeussen V;Coppens J;Koopmans M;Molenkamp R
通讯作者：
Molenkamp R
DOI：
10.1016/j.cmi.2020.09.057
发表时间：
2021-03
期刊：
Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases
影响因子：
--
作者：
Diao B;Wen K;Zhang J;Chen J;Han C;Chen Y;Wang S;Deng G;Zhou H;Wu Y
通讯作者：
Wu Y
影响因子：
38.2
作者：
Chu, Hin;Chan, Jasper Fuk-Woo;Yuen, Kwok-Yung
通讯作者：
Yuen, Kwok-Yung
影响因子：
56.3
作者：
Petersen, Eskild;Koopmans, Marion;Simonsen, Lone
通讯作者：
Simonsen, Lone
影响因子：
4.6
作者：
Balabdaoui F;Mohr D
通讯作者：
Mohr D