Quantitative Detection and Viral Load Analysis of SARS-CoV-2 in Infected Patients
Quantitative Detection and Viral Load Analysis of SARS-CoV-2 in Infected Patients
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DOI：
10.1093/cid/ciaa345
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发表时间：
2020-08-01
影响因子：
11.8
通讯作者：
Zhang, Fujie
中科院分区：
文献类型：
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作者：
Yu, Fengting;Yan, Liting;Zhang, Fujie
背景。 2019 年冠状病毒病 (COVID-19) 已成为突发公共卫生事件。广泛使用的逆转录聚合酶链反应（RT-PCR）方法对于临床诊断和治疗存在局限性。通过液滴数字 PCR (ddPCR) 和基于 2 个目标基因（ORF1ab 和 N）的 RT-PCR 对来自 76 名 COVID-19 确诊患者的总共 323 个样本进行了分析。收集鼻拭子、咽拭子、痰液、血液和尿液。获得临床和影像数据用于临床分期。结果。在两种方法均检测出阳性的 95 个样本中，RT-PCR 的循环阈值 (Ct) 与 ddPCR 的拷贝数（ORF1ab 基因，R-2 = 0.83；N 基因，R-2 = 0.87）高度相关。 RT-PCR 检测的 4 个（4/161）阴性样本和 41 个（41/67）单基因阳性样本根据 ddPCR 检测呈阳性，病毒载量范围为 11.1 至 123.2 拷贝/检测。然后比较呼吸道样本的病毒载量，发现痰中的平均病毒载量（17 429 +/- 6920 拷贝/测试）显着高于咽拭子（2552 +/- 1965 拷贝/测试，P
Background. Coronavirus disease 2019 (COVID-19) has become a public health emergency. The widely used reverse transcription-polymerase chain reaction (RT-PCR) method has limitations for clinical diagnosis and treatment.Methods. A total of 323 samples from 76 COVID-19-confirmed patients were analyzed by droplet digital PCR (ddPCR) and RT-PCR based 2 target genes (ORF1ab and N). Nasal swabs, throat swabs, sputum, blood, and urine were collected. Clinical and imaging data were obtained for clinical staging.Results. In 95 samples that tested positive by both methods, the cycle threshold (Ct) of RT-PCR was highly correlated with the copy number of ddPCR (ORF1ab gene, R-2 = 0.83; N gene, R-2 = 0.87). Four (4/161) negative and 41 (41/67) single-gene positive samples tested by RT-PCR were positive according to ddPCR with viral loads ranging from 11.1 to 123.2 copies/test. The viral load of respiratory samples was then compared and the average viral load in sputum (17 429 +/- 6920 copies/test) was found to be significantly higher than in throat swabs (2552 +/- 1965 copies/test, P