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Analytical Sensitivity and Specificity of Two RT-qPCR Protocols for SARS-CoV-2 Detection Performed in an Automated Workflow
Genes ( IF 2.8 ) Pub Date : 2020-10-12 , DOI: 10.3390/genes11101183
Gustavo Barcelos Barra , Ticiane Henriques Santa Rita , Pedro Góes Mesquita , Rafael Henriques Jácomo , Lídia Freire Abdalla Nery

WHO declared the novel coronavirus (COVID-19) outbreak a global pandemic on 11 March 2020. The establishment of standardized RT-qPCR protocols for respiratory secretions testing, as well as sharing of specimens, data, and information became critical. Here, we investigate the analytical performance of two interim RT-qPCR protocols (Charité and Centers for Disease Control (CDC)) for the qualitative detection of SARS-CoV-2 executed in a fully automated platform. Analytical specificity, PCR amplification efficiency, analytical sensitivity (limit of detection), and cross-reactivity were evaluated using contrived samples. The on-going accuracy was evaluated by retrospective analysis of our test results database (real clinical samples). N1, E, and a modified version of RdRP assays presented adequate analytical specificity, amplification efficiency, and analytical sensitivity using contrived samples. The three assays were applied to all individuals who requested the SARS-CoV-2 molecular test assay in our laboratory and it was observed that N1 gave more positive results than E, and E gave more positive results than RdRP (modified). The RdRP and E were removed from the test and its final version, based on N1 assay only, was applied to 30,699 Brazilian individuals (from 19 February 2020 to 8 May 2020). The aggregated test results available in the database were also presented.

中文翻译:

在自动化工作流程中进行的用于 SARS-CoV-2 检测的两种 RT-qPCR 协​​议的分析灵敏度和特异性

世卫组织于 2020 年 3 月 11 日宣布新型冠状病毒 (COVID-19) 爆发为全球大流行。为呼吸道分泌物检测制定标准化的 RT-qPCR 协​​议以及共享标本、数据和信息变得至关重要。在这里,我们研究了两个临时 RT-qPCR 协​​议(Charité 和疾病控制中心 (CDC))的分析性能,用于在全自动平台上执行的 SARS-CoV-2 定性检测。分析特异性、PCR 扩增效率、分析灵敏度(检测限)和交叉反应性使用人造样品进行评估。通过对我们的测试结果数据库(真实临床样本)的回顾性分析来评估持续的准确性。N1、E 和 RdRP 检测的改进版本提供了足够的分析特异性、扩增效率、和使用人造样品的分析灵敏度。这三种检测适用于我们实验室要求进行 SARS-CoV-2 分子检测检测的所有个体,观察到 N1 的阳性结果多于 E,而 E 的阳性结果多于 RdRP(修改后)。RdRP 和 E 从测试中删除,其最终版本仅基于 N1 检测,应用于 30,699 名巴西人(2020 年 2 月 19 日至 2020 年 5 月 8 日)。还提供了数据库中可用的汇总测试结果。RdRP 和 E 从测试中删除,其最终版本仅基于 N1 检测,应用于 30,699 名巴西人(2020 年 2 月 19 日至 2020 年 5 月 8 日)。还提供了数据库中可用的汇总测试结果。RdRP 和 E 从测试中删除,其最终版本仅基于 N1 检测,应用于 30,699 名巴西人(2020 年 2 月 19 日至 2020 年 5 月 8 日)。还提供了数据库中可用的汇总测试结果。
更新日期:2020-10-12
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