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客户采用我司免疫磁珠分离外泌体发文在Talanta

2021-11-8 22:55:50点击:
Qiuyuan Lin, Zhipeng Huang, Xin Ye, Bin Yang, Xueen Fang, Baohong Liu, Hui Chen, Jilie Kong,
Lab in a tube: Isolation, extraction, and isothermal amplification detection of exosomal long noncoding RNA of gastric cancer,
Talanta,
Volume 225,
2021,
122090,
ISSN 0039-9140,
https://doi.org/10.1016/j.talanta.2021.122090.
(https://www.sciencedirect.com/science/article/pii/S0039914021000114)
Abstract: 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.

Keywords: Exosomal long noncoding RNA; Immunomagnetic beads isolation; Nucleic acid extraction; Loop-mediated isothermal amplification (LAMP)

摘要:肿瘤来源的外泌体继承了亲代细胞的分子信息,在癌症诊断方面具有广阔的前景。目前,两个主要的技术挑战,费时费力的外泌体分离和回收率有限的核酸提取限制了超低丰度外泌体核酸的检测。在这里,我们提出了一种用于检测外泌体核酸的简单、高效和“管中实验室”系统,该系统完全整合了使用免疫磁珠(IMB)(10 分钟)的外泌体富集,基于 NP-40 裂解物的快速外泌体裂解( 5 分钟)和灵敏的环介导等温扩增 (LAMP) 在管中。该方法通过检测胃癌的两种外泌体长非编码RNA生物标志物(HOTTIP和lncRNA-GC1)来证明,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品中的外泌体 HOTTIP RNA 方面表现出良好的性能,具有检测癌症中低丰度外泌体核酸生物标志物的潜力。
关键词:外泌体长链非编码RNA;免疫磁珠分离;核酸提取;环介导等温扩增 (LAMP)