评估多重数字液滴 RT-PCR 作为鼻咽拭子和唾液样本中 SARS-CoV-2 检测的诊断工具。
Assessment of Multiplex Digital Droplet RT-PCR as a Diagnostic Tool for SARS-CoV-2 Detection in Nasopharyngeal Swabs and Saliva Samples.
机构信息
Department of Genetics, Rouen University Hospital and Inserm U1245, UNIROUEN, Normandie University, Normandy Center for Genomic and Personalized Medicine, Rouen, France.
Virology Laboratory, Rouen University Hospital, Rouen, France.
出版信息
Clin Chem. 2021 Apr 29;67(5):736-741. doi: 10.1093/clinchem/hvaa323.
BACKGROUND
Reverse transcription-quantitative PCR on nasopharyngeal swabs is currently the reference COVID-19 diagnosis method but exhibits imperfect sensitivity.
METHODS
We developed a multiplex reverse transcription-digital droplet PCR (RT-ddPCR) assay, targeting 6 SARS-CoV-2 genomic regions, and evaluated it on nasopharyngeal swabs and saliva samples collected from 130 COVID-19 positive or negative ambulatory individuals, who presented symptoms suggestive of mild or moderate SARS-CoV2 infection.
RESULTS
For the nasopharyngeal swab samples, the results obtained using the 6-plex RT-ddPCR and RT-qPCR assays were all concordant. The 6-plex RT-ddPCR assay was more sensitive than RT-qPCR (85% versus 62%) on saliva samples from patients with positive nasopharyngeal swabs.
CONCLUSION
Multiplex RT-ddPCR represents an alternative and complementary tool for the diagnosis of COVID-19, in particular to control RT-qPCR ambiguous results. It can also be applied to saliva for repetitive sampling and testing individuals for whom nasopharyngeal swabbing is not possible.
背景
鼻咽拭子的逆转录定量 PCR 目前是 COVID-19 诊断的参考方法，但灵敏度不理想。
方法
我们开发了一种针对 6 个 SARS-CoV-2 基因组区域的多重逆转录数字液滴 PCR（RT-ddPCR）检测方法，并在来自 130 例 COVID-19 阳性或阴性门诊个体的鼻咽拭子和唾液样本上进行了评估，这些个体出现了提示轻度或中度 SARS-CoV2 感染的症状。
结果
对于鼻咽拭子样本，使用 6 重 RT-ddPCR 和 RT-qPCR 检测方法获得的结果均一致。对于来自鼻咽拭子阳性患者的唾液样本，6 重 RT-ddPCR 检测方法比 RT-qPCR 更灵敏（85%比 62%）。
结论
多重 RT-ddPCR 是 COVID-19 诊断的替代和补充工具，特别是可以控制 RT-qPCR 结果的不明确性。它也可以应用于唾液，用于对不能进行鼻咽拭子采样的个体进行重复采样和检测。
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