Massively scaled-up testing for SARS-CoV-2 RNA via next-generation sequencing of pooled and barcoded nasal and saliva samples.
Massively scaled-up testing for SARS-CoV-2 RNA via next-generation sequencing of pooled and barcoded nasal and saliva samples.
复制标题
DOI：
10.1038/s41551-021-00754-5
复制
发表时间：
2021-07
影响因子：
28.1
通讯作者：
中科院分区：
文献类型：
--
作者：
对无症状人群进行频繁和广泛的检测对于减少严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的不受控制的传播至关重要。尽管检测能力有所提高，但定量聚合酶链式反应检测不能轻易部署到全人群筛查工作所需的规模。在这里，我们报告了一项针对SARS-CoV-2的高通量测试，该测试使用了标记有样本特定分子条形码的下一代混合样本测序。这项测试不需要提取RNA，可以在一次运行中同时分析数千个鼻腔或唾液样本。该检测采用了体外RNA标准，该标准提高了定量的准确性，并需要最低限度的样品到样品的归一化，从而促进了真阴性的调用。我们的实验室在不到两个月的时间里进行了80,000多次测试，证实了大规模提供廉价的结果，具有高度的敏感性和特异性，周转时间不到24小时。这项检测可以迅速调整，以瞄准其他病原体的传播。用于高通量筛选临床样本的临床流水线，通过样本特定的分子条形码汇集在单个下一代测序运行中。
Frequent and widespread testing of the asymptomatic population is essential to mitigate the uncontrolled transmission of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Despite increases in testing capacity, quantitative polymerase chain reaction tests cannot be easily deployed at the scale required for population-wide screening efforts. Here, we report a high-throughput test for SARS-CoV-2 that uses next-generation sequencing of pooled samples tagged with sample-specific molecular barcodes. This test does not require RNA extraction and can simultaneously analyze thousands of nasal or saliva samples in a single run. The test incorporates an in vitro RNA standard which improves quantitation accuracy and requires minimal sample-to-sample normalization, thus facilitating the calling of true negatives. Our laboratory performed over 80,000 tests in under two months, confirming the inexpensive provision of results at scale, with high sensitivity and specificity, and with turn-around times of less than 24 hours. The test can be rapidly adapted to target the spread of other pathogens. A clinical pipeline for high-throughput screening of clinical samples, pooled in a single next-generation sequencing run, via sample-specific molecular barcoding.
登录
查看更多内容
影响因子：
5.8
作者：
Morgan, Martin;Anders, Simon;Gentleman, Robert
通讯作者：
Gentleman, Robert
影响因子：
11.8
作者：
Furukawa, Nathan W;Brooks, John T;Sobel, Jeremy
通讯作者：
Sobel, Jeremy
影响因子：
5.9
作者：
Cheng, Cecilia;Wong, Wai-man;Tsang, Kenneth W.
通讯作者：
Tsang, Kenneth W.
影响因子：
--
作者：
Quail, Michael A;Swerdlow, Harold;Turner, Daniel J
通讯作者：
Turner, Daniel J
影响因子：
33.9
作者：
Kimball, Anne;Hatfield, Kelly M.;Jernigan, John A.
通讯作者：
Jernigan, John A.