Department of Clinical Laboratory, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, 400038, China.
Department of Pathology, 922th Hospital of the People's Liberation Army, Hengyang, 421002, Hunan, China.
Anal Bioanal Chem. 2020 Jan;412(3):601-609. doi: 10.1007/s00216-019-02211-4. Epub 2020 Jan 2.
Numerous studies have shown that exosomes are closely related to the pathogenesis of various diseases, especially cancers. Therefore, a rapid and sensitive method for exosome detection will be of great importance for the diagnosis and prognosis of diseases. We report here a method for exosome detection based on the CD63 aptamer and clustered regular interspaced short palindromic repeats (CRISPR)/Cas12a system. This method consists mainly of exosomal membrane protein recognition based on the CD63 aptamer and signal amplification based on CRISPR/Cas12a. The CD63 aptamer, as an easily adaptable nucleic acid strand, is responsible for the conversion of the amounts of exosomes into nucleic acid detection, whereas CRISPR/Cas12a is responsible for highly specific nucleic acid signal amplification. The detection range of the method was determined as 3 × 10-6 × 10 particles per microliter. Additionally, we successfully applied this method to detect exosomes in clinical samples from both healthy individuals and patients with lung cancer, and the results were highly consistent with those obtained by nanoparticle tracking analysis. In general, this method provides a highly sensitive and specific method for the detection of exosomes and offers an avenue toward future exosome-based diagnosis of diseases.
许多研究表明，外泌体与各种疾病（尤其是癌症）的发病机制密切相关。因此，一种快速、灵敏的外泌体检测方法对于疾病的诊断和预后将具有重要意义。我们在这里报告了一种基于 CD63 适体和簇状规则间隔短回文重复序列（CRISPR）/Cas12a 系统的外泌体检测方法。该方法主要包括基于 CD63 适体的外泌体膜蛋白识别和基于 CRISPR/Cas12a 的信号放大。CD63 适体作为一种易于适配的核酸链，负责将外泌体的数量转换为核酸检测，而 CRISPR/Cas12a 则负责高度特异性的核酸信号放大。该方法的检测范围确定为 3×10-6×10 个/微升。此外，我们成功地将该方法应用于检测来自健康个体和肺癌患者的临床样本中的外泌体，并且结果与纳米颗粒跟踪分析得到的结果高度一致。总的来说，该方法为外泌体的检测提供了一种高度敏感和特异的方法，并为基于外泌体的疾病诊断提供了一种途径。
