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.

中文翻译：

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仿生病毒样颗粒作为严重急性呼吸系统综合症冠状病毒 2 的诊断工具

2019 年冠状病毒病 (COVID-19) 是一种高度传播的疾病，已影响到全球 90% 以上的国家。至少有 1700 万人被感染，一些国家仍在与第一波或第二波疫情作斗争。核酸检测，尤其是逆转录聚合酶链反应 (RT-PCR)，已成为早期检测 COVID-19 感染的主要手段。已经开发出分子测定的阳性对照来验证每个测试并提供高精度。然而，大多数可用的阳性对照品需要冷链配送，不能作为全过程对照。为了克服这些缺点，我们报告了仿生病毒样颗粒（VLP）的生产作为 SARS-CoV-2 阳性对照。用于 RT-PCR 的 SARS-CoV-2 检测模块被封装到来自噬菌体和植物病毒的 VLP 中。嵌合VLP通过目标检测模块和外壳蛋白的体内重建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。这些基于 VLP 的阳性对照模仿 SARS-CoV-2 包装的核糖核酸 (RNA)，但不具有感染性。最重要的是，我们证明阳性对照是可扩展的、稳定的，并且可以广泛用作从 RNA 提取到临床环境中 PCR 的对照。