Department of Biological Sciences, Olivet Nazarene Universitygrid.261375.4, Bourbonnais, Illinois, USA.
Microbiol Spectr. 2021 Oct 31;9(2):e0108921. doi: 10.1128/Spectrum.01089-21. Epub 2021 Oct 13.
Routine testing for SARS-CoV-2 is rare for institutes of higher education due to prohibitive costs and supply chain delays. During spring 2021, we routinely tested all residential students 1 to 2 times per week using pooled, RNA-extraction-free, reverse transcription quantitative PCR (RT-qPCR) testing of saliva at a cost of $0.43/sample with same-day results. The limit of detection was 500 copies/ml on individual samples, and analysis indicates 1,000 and 2,500 copies/ml in pools of 5 and 10, respectively, which is orders of magnitude more sensitive than rapid antigen tests. Importantly, saliva testing flagged 83% of semester positives (43,884 tests administered) and was 95.6% concordant with nasopharyngeal diagnostic results (69.0% concordant on the first test when the nucleocapsid gene (N1) cycle threshold () value was >30). Moreover, testing reduced weekly cases by 59.9% in the spring despite far looser restrictions, allowing for more normalcy while eliminating outbreaks. We also coupled our testing with a survey to clarify symptoms and transmissibility among college-age students. While only 8.5% remained asymptomatic throughout, symptoms were disparate and often cold-like (e.g., only 37.3% developed a fever), highlighting the difficulty with relying on symptom monitoring among this demographic. Based on reported symptom progression, we estimate that we removed 348 days of infectious individuals by routine testing. Interestingly, viral load ( value) at the time of testing did not affect transmissibility ( = 0.0085), though those experiencing noticeable symptoms at the time of testing were more likely to spread the virus to close contacts (31.6% versus 14.3%). Together, our findings support routine testing for reducing the spread of SARS-CoV-2. Implementation of cost- and resource-efficient approaches should receive strong consideration in communities that lack herd immunity. This study highlights the utility of routine testing for SARS-CoV-2 using pooled saliva while maintaining high sensitivity of detection (under 2,500 copies/ml) and rapid turnaround of high volume (up to 930 samples in 8 h by two technicians and one quantitative PCR [qPCR] machine). This pooled approach allowed us to test all residential students 1 to 2 times per week on our college campus during the spring of 2021 and flagged 83% of our semester positives. Most students were asymptomatic or presented with symptoms mirroring common colds at the time of testing, allowing for removal of infectious individuals before they otherwise would have sought testing. To our knowledge, the total per-sample consumable cost of $0.43 is the lowest to date. With many communities still lagging in vaccination rates, routine testing that is cost-efficient highlights the capacity of the laboratory's role in controlling the spread of SARS-CoV-2.
由于成本高昂和供应链延迟,高等院校很少对 SARS-CoV-2 进行常规检测。在 2021 年春季,我们使用无 RNA 提取、逆转录定量 PCR(RT-qPCR)对唾液进行每周 1 到 2 次的 pooled 检测,每次检测的成本为 0.43 美元/样本,当天即可得出结果。单个样本的检测下限为 500 拷贝/ml,而在 5 份和 10 份的 pooled 中分别为 1000 和 2500 拷贝/ml,这比快速抗原检测灵敏几个数量级。重要的是,唾液检测标记了 83%的学期阳性病例(共进行了 43884 次检测),与鼻咽诊断结果的符合率为 95.6%(当核衣壳基因(N1)的循环阈值(Ct)值>30 时,第一次检测的符合率为 69.0%)。此外,尽管限制措施要宽松得多,但检测在春季每周仍减少了 59.9%的病例,从而允许更多的正常活动,同时消除了疫情爆发。我们还将检测与一项调查相结合,以明确大学生群体中的症状和传染性。虽然只有 8.5%的人始终无症状,但症状各不相同,且通常类似于感冒(例如,只有 37.3%的人发烧),这突出表明,在这一年龄段,依靠症状监测存在困难。根据报告的症状进展,我们估计通过常规检测消除了 348 天的传染性个体。有趣的是,检测时的病毒载量(Ct 值)并不影响传染性(=0.0085),尽管那些在检测时出现明显症状的个体更有可能将病毒传播给密切接触者(31.6%比 14.3%)。总的来说,我们的研究结果支持通过常规检测来减少 SARS-CoV-2 的传播。在缺乏群体免疫的社区中,应大力考虑实施成本效益高的方法。本研究强调了使用 pooled 唾液进行 SARS-CoV-2 常规检测的实用性,同时保持了高检测灵敏度(低于 2500 拷贝/ml)和快速高容量(多达 930 个样本在 8 小时内由两名技术人员和一台 qPCR 机器完成)。这种 pooled 方法使我们能够在 2021 年春季每周对我们大学校园的所有住校学生进行 1 到 2 次检测,并标记了我们学期阳性病例的 83%。大多数学生在检测时无症状,或表现出类似于感冒的症状,从而在他们寻求检测之前,就将感染个体隔离了。据我们所知,每个样本的总消耗成本为 0.43 美元,是迄今为止最低的。由于许多社区的疫苗接种率仍然滞后,成本效益高的常规检测突显了实验室在控制 SARS-CoV-2 传播方面的作用。