Department of Chemistry & State Key Laboratory of Molecular Engineering of Polymers, Fudan University, Shanghai, 200433, China; Human Phenome Institute, State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, 200438, China.
Department of Chemistry & State Key Laboratory of Molecular Engineering of Polymers, Fudan University, Shanghai, 200433, China.
Anal Chim Acta. 2022 Jun 15;1212:339937. doi: 10.1016/j.aca.2022.339937. Epub 2022 May 13.
Until now, corona virus disease 2019 (COVID-19) remained to be an enormous threat for global health. As one viral illness induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), versatile, rapid and sensitive method for SARS-CoV-2 detection in early stage is urgently needed. Here, we reported an ultrasensitive and visual in-one-tube detection method which could be accomplished within half an hour from sampling-to-result. By integrating all reactions in one tube, liquid handling steps were omitted and amplicon contamination could be totally avoided. Magnetic beads were employed to achieve the fast extraction of viral nucleic acid and increase the sensitivity. Using portable thermocycler and blue light, the fluorescent results could be directly observed by naked eyes. The proposed method is of higher specificity and sensitivity, nearly at single molecule level. More important, results demonstrated 100% positive detection rate for 40 clinical samples, which was consistent with standard RT-PCR. Thus, our method is considerably simple, rapid, sensitive and accurate, holding great promise for the instant detecting of viruses including SARS-CoV-2 and the next generation of molecular diagnosis.
截至目前，2019 年冠状病毒病 (COVID-19) 仍然对全球健康构成巨大威胁。作为一种由严重急性呼吸系统综合征冠状病毒 2 (SARS-CoV-2) 引起的病毒性疾病，迫切需要一种用于早期 SARS-CoV-2 检测的多功能、快速和敏感的方法。在这里，我们报道了一种超灵敏和可视化的一体式检测方法，从采样到结果可在半小时内完成。通过将所有反应整合在一个管中，省去了移液步骤，可完全避免扩增子污染。采用磁珠实现病毒核酸的快速提取，提高了灵敏度。使用便携式热循环仪和蓝光，可通过肉眼直接观察荧光结果。该方法具有更高的特异性和灵敏度，几乎达到单分子水平。更重要的是，结果表明，对于 40 份临床样本的检测，阳性检出率为 100%，与标准 RT-PCR 一致。因此，我们的方法简便、快速、灵敏、准确，有望用于即时检测包括 SARS-CoV-2 在内的病毒，以及下一代分子诊断。
