Xu Jingsong, Wang Xi, Yang Shuang, He Lei, Wang Yuting, Li Jiajun, Liu Qian, Li Min, Wang Hua
Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Front Microbiol. 2023 Jan 12;13:1070831. doi: 10.3389/fmicb.2022.1070831. eCollection 2022.
The coronavirus disease 2019 (COVID-19) caused by novel severe acute respiratory coronavirus 2 (SARS-CoV-2) has been rapidly spreading worldwide. Rapid and widespread testing is essential to promote early intervention and curb the ongoing COVID-19 pandemic. Current gold standard reverse transcription-polymerase chain reaction (RT-PCR) for detecting SARS-CoV-2 is restricted to professional laboratories and well-trained personnel, thus, limiting its widespread use in resource-limited conditions. To overcome these challenges, we developed a rapid and convenient assay using a versatile integrated tube for the rapid and visual detection of SARS-CoV-2. The reaction conditions of the method were optimized using SARS-CoV-2 RNA standards and the sensitivity and specificity were further determined. Finally, it was verified on clinical specimens. The assay was completed within 40 min, and the result was visible by the naked eye. The limits of detection (LODs) for the target ORF1ab and N genes were 50 copies/μl. No cross-reactivity was observed with the RNA standard samples of four respiratory viruses or clinical samples of common respiratory viral infections. Ninety SARS-CoV-2 positive and 30 SARS-CoV-2 negative patient specimens were analyzed. We compared these results to both prior and concurrent RT-PCR evaluations. As a result, the overall sensitivity and specificity for detection SARS-CoV-2 were 94.5 and 100.0%, respectively.
The integrated tube assay has the potential to provide a simple, specific, sensitive, one-pot, and single-step assay for SARS-CoV-2.
由新型严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的2019冠状病毒病（COVID-19）已在全球迅速传播。快速且广泛的检测对于促进早期干预和遏制当前的COVID-19大流行至关重要。目前用于检测SARS-CoV-2的金标准逆转录聚合酶链反应（RT-PCR）仅限于专业实验室和训练有素的人员，因此限制了其在资源有限条件下的广泛应用。为克服这些挑战，我们开发了一种使用通用集成管的快速便捷检测方法，用于快速可视化检测SARS-CoV-2。使用SARS-CoV-2 RNA标准优化了该方法的反应条件，并进一步确定了其灵敏度和特异性。最后，在临床标本上进行了验证。该检测在40分钟内完成，结果肉眼可见。目标ORF1ab和N基因的检测限（LOD）为50拷贝/微升。未观察到与四种呼吸道病毒的RNA标准样品或常见呼吸道病毒感染的临床样品发生交叉反应。分析了90份SARS-CoV-2阳性和30份SARS-CoV-2阴性患者标本。我们将这些结果与之前和同时进行的RT-PCR评估结果进行了比较。结果，检测SARS-CoV-2的总体灵敏度和特异性分别为94.5%和100.0%。
集成管检测法有可能为SARS-CoV-2提供一种简单、特异、灵敏、一锅法和单步检测方法。
