Charles E Schmidt College of Medicine, Florida Atlantic University, Boca Raton, Florida, United States of America.
PLoS One. 2020 Dec 18;15(12):e0244271. doi: 10.1371/journal.pone.0244271. eCollection 2020.
Widespread testing is required to limit the current public health crisis caused by the COVID-19 pandemic. Multiple tests protocols have been authorized by the food and drugs administration (FDA) under an emergency use authorization (EUA). The majority of these protocols are based on the gold-standard RT-qPCR test pioneered by the U.S. Centers for Disease Control and Prevention (CDC). However, there is still a widespread lack of testing in the US and many of the clinical diagnostics protocols require extensive human labor and materials that could face supply shortages and present biosafety concerns. Given the need to develop alternative reagents and approaches to provide nucleic-acid testing in the face of heightened demand and potential shortages, we have developed a simplified SARS-CoV-2 testing protocol adapted for its use in research laboratories with minimal molecular biology equipment and expertise. The protocol utilizes TRIzol to purify the viral RNA from different types of clinical specimens, requires minimal BSL-1 precautions and, given its high sensitivity, can be easily adapted to pooling samples strategies.
广泛的检测对于限制当前由 COVID-19 大流行引起的公共卫生危机至关重要。食品和药物管理局 (FDA) 根据紧急使用授权 (EUA) 批准了多种检测方案。这些方案中的大多数都是基于美国疾病控制与预防中心 (CDC) 首创的金标准 RT-qPCR 检测。然而，美国仍然普遍缺乏检测，许多临床诊断检测方案需要大量的人力和材料，这些材料可能面临供应短缺和生物安全问题。鉴于需要开发替代试剂和方法来提供核酸检测，以应对需求的增加和潜在的短缺，我们开发了一种简化的 SARS-CoV-2 检测方案，该方案适用于具有最少分子生物学设备和专业知识的研究实验室。该方案利用 TRIzol 从不同类型的临床标本中纯化病毒 RNA，需要最少的 BSL-1 预防措施，并且由于其高灵敏度，可以很容易地适应样本混合策略。
