Centro de Tecnologia em Vacinas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, CEP: 31210-260, Brazil.
Instituto René Rachou, Fundação Oswaldo Cruz/FIOCRUZ, Belo Horizonte, Minas Gerais, CEP: 30190-002, Brazil.
Braz J Microbiol. 2021 Jun;52(2):531-539. doi: 10.1007/s42770-021-00469-4. Epub 2021 Mar 31.
Accurate testing to detect SARS-CoV-2 RNA is key to counteract the virus spread. Nonetheless, the number of diagnostic laboratories able to perform qPCR tests is limited, particularly in developing countries. We describe the use of a virus-inactivating, denaturing solution (DS) to decrease virus infectivity in clinical specimens without affecting RNA integrity. Swab samples were collected from infected patients and from laboratory personnel using a commercially available viral transport solution and the in-house DS. Samples were tested by RT-qPCR, and exposure to infective viruses was also accessed by ELISA. The DS used did not interfere with viral genome detection and was able to maintain RNA integrity for up to 16 days at room temperature. Furthermore, virus loaded onto DS were inactivated, as attested by attempts to grow SARS-CoV-2 in cell monolayers after DS desalt filtration to remove toxic residues. The DS described here provides a strategy to maintain diagnostic accuracy and protects diagnostic laboratory personnel from accidental infection, as it has helped to protect our lab crew.
准确的检测来检测 SARS-CoV-2 RNA 是对抗病毒传播的关键。尽管如此,能够进行 qPCR 检测的诊断实验室数量有限,特别是在发展中国家。我们描述了一种使用病毒灭活、变性溶液(DS)的方法,在不影响 RNA 完整性的情况下降低临床标本中的病毒感染力。拭子样本取自感染患者和实验室人员,使用市售的病毒运输溶液和内部 DS。通过 RT-qPCR 进行测试,并通过 ELISA 评估感染性病毒的暴露情况。所用的 DS 不会干扰病毒基因组的检测,并且能够在室温下保持 RNA 完整性长达 16 天。此外,加载到 DS 上的病毒被灭活,这可以通过在 DS 脱盐过滤以去除有毒残留物后尝试在细胞单层中培养 SARS-CoV-2 来证明。本文所述的 DS 提供了一种保持诊断准确性的策略,并保护诊断实验室人员免受意外感染,因为它有助于保护我们的实验室人员。