Lin Qiuyuan, Huang Zhipeng, Ye Xin, Yang Bin, Fang Xueen, Liu Baohong, Chen Hui, Kong Jilie
Department of Chemistry, Fudan University, Shanghai, 200438, PR China.
Department of Chemistry, Fudan University, Shanghai, 200438, PR China; Shanghai Stomatological Hospital, Shanghai, 200438, PR China.
Talanta. 2021 Apr 1;225:122090. doi: 10.1016/j.talanta.2021.122090. Epub 2021 Jan 7.
Tumor-derived exosomes that inherit molecular information on parental cells hold great promise for cancer diagnostics. Currently, two main technical challenges, time-consuming and labor-intensive isolation of exosome and nucleic acid extraction with limited recovery that have restricted the detection of ultralow abundance exosomal nucleic acids. Here, we proposed a simple, efficient and "lab in a tube" system for the detection of exosomal nucleic acids, which fully integrated exosomes enrichment using immunomagnetic beads (IMB) (10 min), fast exosomes lysis based on NP-40 lysate (5 min) and sensitive loop-mediated isothermal amplification (LAMP) in a tube. This method was demonstrated by detecting two exosomal long noncoding RNA biomarkers of gastric cancer (HOTTIP and lncRNA-GC1) with a dynamic detection ranging from 300 ng/μL to 10 ng/μL, and the detection limit of LAMP was 10 ng/μL. Additionally, this platform exhibited good performance in the analysis of exosomal HOTTIP RNA directly in human serum samples, which has the potential for detection of low-abundance exosomal nucleic acid biomarkers from cancers.
继承亲代细胞分子信息的肿瘤来源外泌体在癌症诊断方面具有巨大潜力。目前，两个主要技术挑战限制了超低丰度外泌体核酸的检测，即外泌体分离耗时且费力，以及核酸提取回收率有限。在此，我们提出了一种用于检测外泌体核酸的简单、高效且“管中实验室”系统，该系统完全集成了使用免疫磁珠（IMB）富集外泌体（10分钟）、基于NP - 40裂解液的快速外泌体裂解（5分钟）以及管内灵敏的环介导等温扩增（LAMP）。通过检测胃癌的两种外泌体长链非编码RNA生物标志物（HOTTIP和lncRNA - GC1）验证了该方法，其动态检测范围为300 ng/μL至10 ng/μL，LAMP的检测限为10 ng/μL。此外，该平台在直接分析人血清样本中的外泌体HOTTIP RNA时表现良好，具有检测癌症中低丰度外泌体核酸生物标志物的潜力。
