Simpler and faster Covid-19 testing: Strategies to streamline SARS-CoV-2 molecular assays.
Simpler and faster Covid-19 testing: Strategies to streamline SARS-CoV-2 molecular assays.
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更简单、更快速的Covid-19检测：简化SARS-CoV-2分子检测的策略
DOI：
10.1016/j.ebiom.2021.103236
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发表时间：
2021-03
期刊：
影响因子：
11.1
通讯作者：
Lutz BR
中科院分区：
文献类型：
--
作者：
Panpradist N;Wang Q;Ruth PS;Kotnik JH;Oreskovic AK;Miller A;Stewart SWA;Vrana J;Han PD;Beck IA;Starita LM;Frenkel LM;Lutz BR
检测SARS-CoV-2感染对于治疗、隔离感染者和暴露者以及追踪接触者非常重要。RT-qPCR是灵敏检测SARS-CoV-2 RNA的“金标准”方法，但大多数实验室开发的RT-qPCR检测方法涉及复杂的步骤。在这里，我们的目标是通过简化反应设置，消除RNA提取，并提出降低成本的检测工作流程来简化RT-qPCR检测，从而避免对昂贵的qPCR仪器的需求。开发了一种基于低成本RT-PCR的“试剂盒”，其周转速度比CDC开发的方案更快。我们展示了三种检测工作流程：两种可以部署在实验室中进行不同复杂性的检测，另一种可以简单到可以进行即时检测。使用模拟鼻基质中掺入的SARS-CoV-2 RNA评估分析灵敏度。使用人造人鼻基质（n = 41）和从有呼吸道症状的个体中采集的临床鼻标本（n = 110）评价临床性能。使用纯化的SARS-CoV-2 RNA，冻干RT-PCR的分析灵敏度为10个拷贝/反应，使用模拟鼻基质中的直接裂解物时为20个拷贝/反应。对人工人基质的检测性能评价显示，在≥20个RNA拷贝时，特异性为96.7-100%，灵敏度为100%。与临床标本的标准CDC方案进行头对头比较显示，灵敏度为83.8-94.6%，特异性为96.8-100%。我们发现3.6%的不确定样本（未检出的人类对照），低于标准方案的8.1%。这项初步工作应支持实验室或商业实体开发和扩大新冠病毒检测的可及性。该检测试剂盒的软件指南开发正在进行中，以便在其他环境中实施。美国NIH R 01 AI 140845和西雅图儿童研究所
Detection of SARS-CoV-2 infections is important for treatment, isolation of infected and exposed individuals, and contact tracing. RT-qPCR is the “gold-standard” method to sensitively detect SARS-CoV-2 RNA, but most laboratory-developed RT-qPCR assays involve complex steps. Here, we aimed to simplify RT-qPCR assays by streamlining reaction setup, eliminating RNA extraction, and proposing reduced-cost detection workflows that avoid the need for expensive qPCR instruments. A low-cost RT-PCR based “kit” was developed for faster turnaround than the CDC developed protocol. We demonstrated three detection workflows: two that can be deployed in laboratories conducting assays of variable complexity, and one that could be simple enough for point-of-care. Analytical sensitivity was assessed using SARS-CoV-2 RNA spiked in simulated nasal matrix. Clinical performance was evaluated using contrived human nasal matrix (n = 41) and clinical nasal specimens collected from individuals with respiratory symptoms (n = 110). The analytical sensitivity of the lyophilised RT-PCR was 10 copies/reaction using purified SARS-CoV-2 RNA, and 20 copies/reaction when using direct lysate in simulated nasal matrix. Evaluation of assay performance on contrived human matrix showed 96.7–100% specificity and 100% sensitivity at ≥20 RNA copies. A head-to-head comparison with the standard CDC protocol on clinical specimens showed 83.8–94.6% sensitivity and 96.8–100% specificity. We found 3.6% indeterminate samples (undetected human control), lower than 8.1% with the standard protocol. This preliminary work should support laboratories or commercial entities to develop and expand access to Covid-19 testing. Software guidance development for this assay is ongoing to enable implementation in other settings. USA NIH R01AI140845 and Seattle Children's Research Institute
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影响因子：
16.6
作者：
Smyrlaki I;Ekman M;Lentini A;Rufino de Sousa N;Papanicolaou N;Vondracek M;Aarum J;Safari H;Muradrasoli S;Rothfuchs AG;Albert J;Högberg B;Reinius B
通讯作者：
Reinius B
影响因子：
3.7
作者：
Panpradist N;Beck IA;Chung MH;Kiarie JN;Frenkel LM;Lutz BR
通讯作者：
Lutz BR
影响因子：
8.4
作者：
Li, Lang;He, Jian-an;Gu Dayong
通讯作者：
Gu Dayong
影响因子：
3.7
作者：
Panpradist, Nuttada;Toley, Bhushan J.;Lutz, Barry R.
通讯作者：
Lutz, Barry R.
中国南京市密切接触者中筛查出的24例COVID-19无症状感染者的临床特征
影响因子：
9.1
作者：
Hu, Zhiliang;Song, Ci;Shen, Hongbing
通讯作者：
Shen, Hongbing