Biomimetic Virus-Like Particles as Severe Acute Respiratory Syndrome Coronavirus 2 Diagnostic Tools
Biomimetic Virus-Like Particles as Severe Acute Respiratory Syndrome Coronavirus 2 Diagnostic Tools
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DOI：
10.1021/acsnano.0c08430
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发表时间：
2021-01-26
期刊：
影响因子：
17.1
通讯作者：
Steinmetz, Nicole F.
中科院分区：
文献类型：
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作者：
Chan, Soo Khim;Du, Pinyi;Steinmetz, Nicole F.
2019冠状病毒病（COVID-19）是一种高度传播的疾病，影响了全球90%以上的国家。至少有1 700万人受到感染，一些国家仍在与第一波或第二波大流行作斗争。核酸检测，特别是逆转录聚合酶链反应（RT-PCR），已成为早期检测COVID-19感染的主力。已经开发了分子测定的阳性对照，以验证每个检测并提供高准确度。然而，大多数可用的阳性对照需要冷链配送，不能作为全过程控制。为了克服这些缺点，我们报告了仿生病毒样颗粒（VLP）作为SARS-CoV-2阳性对照的生产。将SARS-CoV-2的RT-PCR检测模块分别构建于噬菌体和植物病毒的VLP中。通过靶检测模块和外壳蛋白的体内重构和共表达或通过纯化的检测模块RNA序列和外壳蛋白的体外组装获得嵌合VLP。这些基于VLP的阳性对照模拟SARS-CoV-2包装的核糖核酸（RNA），但无感染性。最重要的是，我们证明了阳性对照是可扩展的、稳定的，并且可以广泛地用作临床环境中从RNA提取到PCR的对照。
Coronavirus disease 2019 (COVID-19) is a highly transmissible disease that has affected more than 90% of the countries worldwide. At least 17 million individuals have been infected, and some countries are still battling first or second waves of the pandemic. Nucleic acid tests, especially reverse transcription polymerase chain reaction (RT-PCR), have become the workhorse for early detection of COVID-19 infection. Positive controls for the molecular assays have been developed to validate each test and to provide high accuracy. However, most available positive controls require cold-chain distribution and cannot serve as full-process control. To overcome these shortcomings, we report the production of biomimetic virus-like particles (VLPs) as SARS-CoV-2 positive controls. A SARS-CoV-2 detection module for RT-PCR was encapsidated into VLPs from a bacteriophage and a plant virus. The chimeric VLPs were obtained either by in vivo reconstitution and coexpression of the target detection module and coat proteins or by in vitro assembly of purified detection module RNA sequences and coat proteins. These VLP-based positive controls mimic SARS-CoV-2 packaged ribonucleic acid (RNA) while being noninfectious. Most importantly, we demonstrated that the positive controls are scalable, stable, and can serve broadly as controls, from RNA extraction to PCR in clinical settings.