VIR-CRISPR: Visual in-one-tube ultrafast RT-PCR and CRISPR method for instant SARS-CoV-2 detection.
VIR-CRISPR: Visual in-one-tube ultrafast RT-PCR and CRISPR method for instant SARS-CoV-2 detection.
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DOI：
10.1016/j.aca.2022.339937
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发表时间：
2022-06-15
影响因子：
6.2
通讯作者：
Qian, Chunyan
中科院分区：
文献类型：
--
作者：
Wang, Rui;Li, Yongfang;Pang, Yanan;Zhang, Fang;Li, Fuyou;Luo, Shihua;Qian, Chunyan
到目前为止，2019冠状病毒病（COVID-19）仍然是对全球健康的巨大威胁。作为严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）引起的一种病毒性疾病，急需一种通用、快速、灵敏的SARS-CoV-2早期检测方法。在这里，我们报告了一种超灵敏和可视化的单管检测方法，可以在半小时内完成从采样到结果。通过将所有反应集成在一个管中，省略了液体处理步骤，并且可以完全避免扩增子污染。采用磁珠法实现了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，荧光结果可以直接用肉眼观察。该方法具有较高的特异性和灵敏度，接近单分子水平。对40份临床标本的检测结果显示，阳性检出率为100%，与标准RT-PCR检测结果一致。因此，我们的方法是相当简单，快速，灵敏度和准确性，持有巨大的希望，即时检测病毒，包括SARS-CoV-2和下一代的分子诊断。我们提出了一种称为VIR-CRISPR的超灵敏，即时和可视化检测方法，用于即时检测SARS-CoV-2。它将超快RT-PCR和CRISPR集成在一个试管中，具有高特异性和近单分子水平的检测灵敏度，为快速临床诊断带来了巨大的希望。
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. We proposed an ultrasensitive, instant and visual detection method termed as VIR-CRISPR for point-of-care SARS-CoV-2 detection. It integrated ultrafast RT-PCR and CRISPR in-one-tube with high specificity and detection sensitivity at nearly single molecule level, holding great promise for rapid clinical diagnosis.
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DOI：
10.1126/science.aav4294
发表时间：
2018-11-16
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Harrington LB;Burstein D;Chen JS;Paez-Espino D;Ma E;Witte IP;Cofsky JC;Kyrpides NC;Banfield JF;Doudna JA
通讯作者：
Doudna JA
影响因子：
4.7
作者：
Li, Linxian;Li, Shiyuan;Wang, Jin
通讯作者：
Wang, Jin
影响因子：
16.6
作者：
Ding X;Yin K;Li Z;Lalla RV;Ballesteros E;Sfeir MM;Liu C
通讯作者：
Liu C
影响因子：
9.3
作者：
Farrar, Jared S.;Wittwer, Carl T.
通讯作者：
Wittwer, Carl T.
影响因子：
5.7
作者：
Huang, Wei E.;Lim, Boon;Cui, Zhanfeng
通讯作者：
Cui, Zhanfeng