College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China.
CAS Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
Anal Chem. 2021 Apr 6;93(13):5606-5611. doi: 10.1021/acs.analchem.1c00335. Epub 2021 Mar 25.
When dealing with infectious pathogens, the risk of contamination or infection in the process of detecting them is nonnegligible. Separation-free detection will be beneficial in operation and safety. In this work, we proposed a DNAzyme walker for homogeneous and isothermal detection of enterovirus. The DNAzyme is divided into two inactivate subunits. When the subunit-conjugated antibody binds to the target virus, the activity of the DNAzyme recovers as a result of spatial proximity. The walker propels, and the fluorescence recovers. The final fluorescence intensity of the reaction mixture is related to the concentration of the target virus. The detection limit of this proposed method is 6.6 × 10 copies/mL for EV71 and 4.3 × 10 copies/mL for CVB3, respectively. Besides, this method was applied in detection of EV71 in clinical samples with a satisfactory result. The entire experiment is easy to operate, and the proposed method has great potential for practical use.
在处理传染性病原体时，检测过程中污染或感染的风险不可忽视。非分离检测在操作和安全性方面将是有益的。在这项工作中，我们提出了一种用于肠道病毒同种型和等温检测的 DNAzyme 行走者。DNAzyme 分为两个失活亚基。当亚基缀合的抗体与靶病毒结合时，由于空间接近，DNAzyme 的活性恢复。行走者推进，荧光恢复。反应混合物的最终荧光强度与靶病毒的浓度有关。该方法的检测限分别为 EV71 的 6.6×10 拷贝/ml 和 CVB3 的 4.3×10 拷贝/ml。此外，该方法还应用于临床样本中 EV71 的检测，结果令人满意。整个实验操作简单，该方法具有很大的实际应用潜力。
