Proposal of RT-PCR-Based Mass Population Screening for Severe Acute Respiratory Syndrome Coronavirus 2 (Coronavirus Disease 2019)
Proposal of RT-PCR-Based Mass Population Screening for Severe Acute Respiratory Syndrome Coronavirus 2 (Coronavirus Disease 2019)
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DOI：
10.1016/j.jmoldx.2020.07.001
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发表时间：
2020-10-01
影响因子：
4.1
通讯作者：
Kolhe, Ravindra
中科院分区：
文献类型：
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作者：
Sahajpal, Nikhil S.;Mondal, Ashis K.;Kolhe, Ravindra
严重急性呼吸系统综合征冠状病毒2型（SARS-CoV-2）检测在许多国家滞后，原因是检测试剂盒短缺和分析过程瓶颈。这项研究调查了2019年大规模人群冠状病毒病筛查样本合并方法的可行性和准确性。对之前检测过SARS-CoV-2的940份鼻咽拭子样本（934份阴性和6份阳性）进行去识别，并分配随机编号进行分析，共生成94个样本池，每个样本池10份。在96孔板中进行自动RNA提取，然后进行RT-PCR。鉴定阳性样本池，并重新分析单个样本。在94个样本池/威尔斯孔中，4个为阳性[Ct值：N（22.7 - 28.3）、ORF 1ab（23.3 - 27.2）和内部对照（34.4 - 35.4）]。对组成4个样本池的40份样本进行了单独鉴定和再分析; 6份样本为阳性，N基因、ORF 1ab和内部对照的Ct值与其各自的威尔斯孔相当。对具有高Ct值的样本进行了额外的实验，与单个检测方法相比，总体结果显示91.6%的阳性一致率和100%的阴性一致率。因此，在148个反应中检测了940份样本，而在常规筛选中检测了940份反应。样本池策略可以帮助赶上测试需求和最小的周转时间，并促进在实验室用品，提取和PCR试剂盒目前短缺的巨大节省。
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing has lagged in many countries because of test kit shortages and analytical process bottlenecks. This study investigated the feasibility and accuracy of a sample pooling approach for wide-scale population screening for coronavirus disease 2019. A total of 940 nasopharyngeal swab samples (934 negative and 6 positive) previously tested for SARS-CoV-2 were deidentified and assigned random numbers for analysis, and 94 pools of 10 samples each were generated. Automated RNA extraction, followed by RT-PCR, was performed in a 96-wed plate. Positive pools were identified, and the individual samples were reanalyzed. Of the 94 pools/wells, four were positive [Ct values: N (22.7 to 28.3), ORF1ab (23.3 to 27.2), and internal control (34.4 to 35.4)]. The 40 samples comprising the four pools were identified and reanalyzed individually; six samples were positive, with Ct values of N gene, ORF1ab, and internal control comparable to their respective wells. Additional experiments were performed on samples with high Ct values, and overall results showed 91.6% positive and 100% negative agreement compared with individual testing approach. Thus, 940 samples were tested in 148 reactions compared with 940 reactions in routine screening. The sample pooling strategy may help catch up with testing needs and minimal turnaround times and facilitate enormous savings on laboratory supplies, extraction, and PCR kits currently in short supply.