Analytical Sensitivity and Specificity of Two RT-qPCR Protocols for SARS-CoV-2 Detection Performed in an Automated Workflow.
Analytical Sensitivity and Specificity of Two RT-qPCR Protocols for SARS-CoV-2 Detection Performed in an Automated Workflow.
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DOI：
10.3390/genes11101183
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发表时间：
2020-10-12
期刊：
影响因子：
3.5
通讯作者：
Nery LFA
中科院分区：
文献类型：
--
作者：
Barra GB;Santa Rita TH;Mesquita PG;Jácomo RH;Nery LFA
世卫组织于二零二零年三月十一日宣布新型冠状病毒（COVID-19）爆发为全球大流行。建立用于呼吸道分泌物检测的标准化RT-qPCR方案以及共享样本、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方案（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的定性检测。使用人工样本评价分析特异性、PCR扩增效率、分析灵敏度（检测限）和交叉反应性。通过对我们的测试结果数据库（真实的临床样本）进行回顾性分析，评价了持续准确性。N1、E和改良版RdRP检测试剂盒使用人工样本显示了足够的分析特异性、扩增效率和分析灵敏度。这三种检测方法应用于所有要求在我们实验室进行SARS-CoV-2分子检测的个体，观察到N1比E给出更阳性的结果，E比RdRP（改良）给出更阳性的结果。从测试中删除了RdRP和E，其最终版本仅基于N1检测，应用于30，699名巴西人（从2020年2月19日至2020年5月8日）。还介绍了数据库中现有的汇总测试结果。
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.
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影响因子：
14.9
作者：
Ruijter JM;Ramakers C;Hoogaars WM;Karlen Y;Bakker O;van den Hoff MJ;Moorman AF
通讯作者：
Moorman AF
影响因子：
6.8
作者：
Lippi, Giuseppe;Simundic, Ana-Maria;Plebani, Mario
通讯作者：
Plebani, Mario
DOI：
10.1007/978-3-642-03683-5_6
发表时间：
2009-07-22
期刊：
Molecular Biology of the SARS-Coronavirus
影响因子：
--
作者：
Ulferts R;Imbert I;Canard B;Ziebuhr J
通讯作者：
Ziebuhr J
影响因子：
12.3
作者：
Davidson, Andrew D.;Williamson, Maia Kavanagh;Matthews, David A.
通讯作者：
Matthews, David A.
影响因子：
19
作者：
Pfefferle, Susanne;Reucher, Svenja;Luetgehetmann, Marc
通讯作者：
Luetgehetmann, Marc