SARS-CoV-2 Coronavirus Nucleocapsid Antigen-Detecting Half-Strip Lateral Flow Assay Toward the Development of Point of Care Tests Using Commercially Available Reagents
SARS-CoV-2 Coronavirus Nucleocapsid Antigen-Detecting Half-Strip Lateral Flow Assay Toward the Development of Point of Care Tests Using Commercially Available Reagents
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DOI：
10.1021/acs.analchem.0c01975
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发表时间：
2020-08-18
影响因子：
7.4
通讯作者：
Nichols, Kevin P.
中科院分区：
文献类型：
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作者：
Grant, Benjamin D.;Anderson, Caitlin E.;Nichols, Kevin P.
SARS-CoV-2大流行对快速诊断检测产生了前所未有的需求，以有效治疗和缓解COVID-19。目前采用的主要诊断工具是逆转录聚合酶链反应（RT-PCR），它可以具有良好的灵敏度和优异的特异性。不幸的是，在全球SARS-CoV-2大流行期间，试剂的实施成本和物流问题阻碍了其按需普及。侧流检测（LFA）是一类诊断方法，如果临床上足够敏感，可能会填补目前RT-PCR检测制度中的许多空白，特别是在低收入和中等收入国家（LMIC）。迄今为止，已经开发了许多血清学LFA，但没有一种满足诊断用例所需的性能要求，主要是由于感染和血清转化之间的相对较长的延迟。然而，基于先前报道的SARS-CoV-1结果，基于抗原的SARS-CoV-2检测可能比血清学检测具有显著更好的临床灵敏度。迄今为止，仅开发了非常少量的抗原检测LFA。开发半条LFA是开发任何LFA格式的有用的第一步。在这项工作中，我们提出了一个半条LFA使用市售抗体检测SARS-CoV-2。我们在缓冲液中检测了该LFA，并使用灵敏度与目视读数相当的光学读数器测量了重组抗原的LOD为0.65 ng/mL（95% CI为0.53 - 0.77 ng/mL）ng/mL。需要进一步开发，包括评价适当的样品基质，使这种测定方法在护理点环境中有用，尽管这种半条LFA可以作为其他开发类似测试的有用起点。
The SARS-CoV-2 pandemic has created an unprecedented need for rapid diagnostic testing to enable the efficient treatment and mitigation of COVID-19. The primary diagnostic tool currently employed is reverse transcription polymerase chain reaction (RT-PCR), which can have good sensitivity and excellent specificity. Unfortunately, implementation costs and logistical problems with reagents during the global SARS-CoV-2 pandemic have hindered its universal on demand adoption. Lateral flow assays (LFAs) represent a class of diagnostic that, if sufficiently clinically sensitive, may fill many of the gaps in the current RT-PCR testing regime, especially in low- and middle-income countries (LMICs). To date, many serology LFAs have been developed, though none meet the performance requirements necessary for diagnostic use cases, primarily due to the relatively long delay between infection and seroconversion. However, on the basis of previously reported results from SARS-CoV-1, antigen-based SARS-CoV-2 assays may have significantly better clinical sensitivity than serology assays. To date, only a very small number of antigen-detecting LFAs have been developed. Development of a half-strip LFA is a useful first step in the development of any LFA format. In this work, we present a half-strip LFA using commercially available antibodies for the detection of SARS-CoV-2. We have tested this LFA in buffer and measured an LOD of 0.65 ng/mL (95% CI of 0.53 to 0.77 ng/mL) ng/mL with recombinant antigen using an optical reader with sensitivity equivalent to a visual read. Further development, including evaluating the appropriate sample matrix, will be required for this assay approach to be made useful in a point of care setting, though this half-strip LFA may serve as a useful starting point for others developing similar tests.