Rapid and sensitive exosome detection with CRISPR/Cas12a

• Published in: Analytical and bioanalytical chemistry 2020, Vol.412 (3), p.601-609
• Main Authors: Zhao, Xianxian, Zhang, Wenqing, Qiu, Xiaopei, Mei, Qiang, Luo, Yang, Fu, Weiling
• Format: Article
• Language: English
• Subjects: A549 Cells; Amplification; Analytical Chemistry; Aptamers; Aptamers, Nucleotide - chemistry; Biochemistry; Biosensing Techniques - methods; CD63 antigen; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; CRISPR; CRISPR-Cas Systems; Diagnosis; Ethylenediaminetetraacetic acid; Exosomes; Exosomes - chemistry; Exosomes - pathology; Food Science; Humans; Laboratory Medicine; Lung cancer; Lung Neoplasms - pathology; Membrane proteins; Monitoring/Environmental Analysis; Nanoparticles; Nucleic Acid Amplification Techniques - methods; Nucleic acids; Pathogenesis; Prognosis; Research Paper; Tetraspanin 30 - analysis
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-019-02211-4

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 3 –6 × 10 7 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.