Department of Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine, Kyoto, Japan.
Microbiol Spectr. 2024 Mar 5;12(3):e0368423. doi: 10.1128/spectrum.03684-23. Epub 2024 Jan 30.
Pooled testing combined with molecular diagnostics for the detection of SARS-CoV-2 is a promising method that can increase testing capacities and save costs. However, pooled testing is also associated with the risks of decreased test sensitivity and specificity. To perform reliable pooled testing, we developed and validated three automated media pooling and molecular diagnostic systems. These pooling systems (geneLEAD-PS, Panther-PS, and Biomek-PS) comprised existing automated molecular detection platforms, corresponding automated media pooling devices, and laboratory information management systems. Analytical sensitivity analysis and mock sample evaluation were performed, and the obtained data were used to determine the sizes of the pool for the validation study. In the validation study, a total of 2,448, 3,228, and 6,420 upper respiratory samples were used for geneLEAD-PS, Panther-PS, and Biomek-PS, respectively, and the diagnostic performances were compared with the reference RT‒PCR assay. A pool size of 6 for geneLEAD-PS and a pool size of 4 for Panther-PS and Biomek-PS were selected for the validation studies. All three systems showed high positive percent agreement values of ≥90.5% and negative percent agreement values of ≥99.8% for any specimen type. Pooled testing resulted in a 65%-71% reduction in cost per sample. The testing capacities of geneLEAD-PS, Panther-PS, and Biomek-PS were 144 samples in 3 hours, 384 samples in 5.5 hours, and 376 samples in 4 hours, respectively. The developed pooling systems showed robust diagnostic performances and will increase the testing capacities of molecular diagnostic tests while saving costs and may contribute to infection control of COVID-19.IMPORTANCEDuring the COVID-19 pandemic, there have been surges in demand for accurate molecular diagnostic testing and laboratory supply shortages. Pooled testing combined with highly sensitive molecular testing, which entails mixing multiple samples as a single sample, is a promising approach to increase testing capacities while reducing the use of consumables. However, pooled testing is associated with risks that compromise diagnostic performance, such as false negatives due to dilution of positive samples or false positives due to cross-contamination. To perform reliable pooled testing, three different pooling systems (an automated pooling device, an automated molecular detection platform, and a laboratory information management system) were developed to accurately interpret pooled testing results. These three systems were validated using multiple clinical samples and showed high concordance with individual testing. The developed pooling systems will contribute to increasing reliable molecular testing capacities while using fewer consumables and saving costs.
汇总检测结合分子诊断用于检测 SARS-CoV-2 是一种很有前途的方法，可以提高检测能力并节省成本。然而，汇总检测也存在检测灵敏度和特异性降低的风险。为了进行可靠的汇总检测，我们开发并验证了三种自动化介质汇总和分子诊断系统。这些汇总系统（geneLEAD-PS、Panther-PS 和 Biomek-PS）包括现有的自动化分子检测平台、相应的自动化介质汇总设备和实验室信息管理系统。进行了分析灵敏度分析和模拟样本评估，所得数据用于确定验证研究中的池大小。在验证研究中，分别使用了总共 2448、3228 和 6420 个上呼吸道样本进行 geneLEAD-PS、Panther-PS 和 Biomek-PS，将诊断性能与参考 RT-PCR 检测进行了比较。对于 geneLEAD-PS，池大小为 6，对于 Panther-PS 和 Biomek-PS，池大小为 4，用于验证研究。对于任何标本类型，三种系统的阳性百分比一致性值均≥90.5%，阴性百分比一致性值均≥99.8%。汇总检测可使每个样本的成本降低 65%-71%。geneLEAD-PS、Panther-PS 和 Biomek-PS 的检测能力分别为 3 小时内 144 个样本、5.5 小时内 384 个样本和 4 小时内 376 个样本。开发的汇总系统表现出稳健的诊断性能，将提高分子诊断测试的检测能力，同时节省成本，并有助于控制 COVID-19 的感染。
在 COVID-19 大流行期间，对准确的分子诊断测试和实验室供应的需求激增。汇总检测结合高灵敏度的分子检测，即将多个样本混合为单个样本，是一种有前途的增加检测能力的方法，同时减少消耗品的使用。然而，汇总检测存在影响诊断性能的风险，例如阳性样本因稀释而出现假阴性，或因交叉污染而出现假阳性。为了进行可靠的汇总检测，开发了三种不同的汇总系统（自动汇总设备、自动分子检测平台和实验室信息管理系统），以准确解释汇总检测结果。使用多个临床样本对这三种系统进行了验证，结果与单独检测高度一致。开发的汇总系统将有助于在使用更少消耗品和节省成本的情况下，增加可靠的分子检测能力。
