Evaluation of COVID-19 RT-qPCR Test in Multi sample Pools
Evaluation of COVID-19 RT-qPCR Test in Multi sample Pools
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DOI：
10.1093/cid/ciaa531
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发表时间：
2020-10-15
影响因子：
11.8
通讯作者：
Kishony, Roy
中科院分区：
文献类型：
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作者：
Yelin, Idan;Aharony, Noga;Kishony, Roy
背景最近出现的严重急性呼吸综合征冠状病毒2（SARS-CoV-2）导致了目前规模空前的大流行。尽管诊断测试对于检测和响应能力至关重要，但不堪重负的医疗保健系统已经经历了与该测试相关的试剂短缺，需要一种精简的立即适用的方案。使用逆转录定量聚合酶链反应检测阳性样本的RNA提取物是否存在SARS-CoV-2，单独或在不同大小的样本池（2-，4-，8-，16-，32-和64-样本池）中与阴性样本一起使用。另外3份阳性样本的转运培养基与阴性样本的转运培养基在8份样本池中混合时也进行了检测。使用标准试剂盒和方案，可以在多达32个样本的池中检测到单个阳性样本，估计假阴性率为10%。也可以检测稀释至64份的阳性样品，尽管这可能需要额外的扩增循环。在RNA提取后或提取前进行合并时，可检测到单一阳性样本。由于它使用标准方案、试剂和设备，因此这种合并方法可以立即应用于当前的临床检测实验室。我们希望，2019年冠状病毒病合并检测的实施将扩大目前的筛查能力，从而能够扩大社区以及医院部门、军队或工厂轮班等密切有机群体的检测范围。
Background. The recent emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) led to a current pandemic of unprecedented scale. Although diagnostic tests are fundamental to the ability to detect and respond, overwhelmed healthcare systems are already experiencing shortages of reagents associated with this test, calling for a lean immediately applicable protocol.Methods. RNA extracts of positive samples were tested for the presence of SARS-CoV-2 using reverse transcription quantitative polymerase chain reaction, alone or in pools of different sizes (2-, 4-, 8-, 16-, 32-, and 64-sample pools) with negative samples. Transport media of additional 3 positive samples were also tested when mixed with transport media of negative samples in pools of 8.Results. A single positive sample can be detected in pools of up to 32 samples, using the standard kits and protocols, with an estimated false negative rate of 10%. Detection of positive samples diluted in even up to 64 samples may also be attainable, although this may require additional amplification cycles. Single positive samples can be detected when pooling either after or prior to RNA extraction.Conclusions. As it uses the standard protocols, reagents, and equipment, this pooling method can be applied immediately in current clinical testing laboratories. We hope that such implementation of a pool test for coronavirus disease 2019 would allow expanding current screening capacities, thereby enabling the expansion of detection in the community, as well as in close organic groups, such as hospital departments, army units, or factory shifts.