Multiplexed and Extraction-Free Amplification for Simplified SARSCoV-2 RT-PCR Tests
Multiplexed and Extraction-Free Amplification for Simplified SARSCoV-2 RT-PCR Tests
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DOI：
10.1021/acs.analchem.0c03918
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发表时间：
2021-02-25
影响因子：
7.4
通讯作者：
Connelly, John T.
中科院分区：
文献类型：
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作者：
Byrnes, Samantha A.;Gallagher, Ryan;Connelly, John T.
全球COVID-19大流行的迅速爆发导致准确诊断疾病的挑战，包括样本采集、保存和纯化的供应短缺。目前，大多数诊断测试需要RNA提取和RT-PCR检测;然而，提取是昂贵和耗时的，并且需要技术专长。考虑到这些挑战，我们报告了来自246个临床样本的SARSCoV-2 RNA的无提取多重扩增，灵敏度为86%，特异性为100%。多重RT-PCR使用CDC单重靶标，LoD为2 c/mu L。我们还报告了在不同的运输介质中使用一系列主混合物的扩增。这项工作可以帮助指导在整个大流行期间，当供应品的可用性发生变化时，哪些试剂组合将能够获得准确的结果。实施这些方法可以降低复杂性和成本，最大限度地减少试剂的使用，加快结果的时间，并增加测试能力。
The rapid onset of the global COVID-19 pandemic has led to challenges for accurately diagnosing the disease, including supply shortages for sample collection, preservation, and purification. Currently, most diagnostic tests require RNA extraction and detection by RT-PCR; however, extraction is expensive and time-consuming and requires technical expertise. With these challenges in mind, we report extraction-free, multiplexed amplification of SARSCoV-2 RNA from 246 clinical samples, resulting in 86% sensitivity and 100% specificity. The multiplex RT-PCR uses the CDC singleplex targets and has an LoD of 2 c/mu L. We also report on amplification using a range of master mixes in different transport media. This work can help guide which combinations of reagents will enable accurate results when availability of supplies changes throughout the pandemic. Implementing these methods can reduce complexity and cost, minimize reagent usage, expedite time to results, and increase testing capacity.