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CRISPR/Cas13a-Powered Electrochemical Microfluidic Biosensor for Nucleic Acid Amplification-Free miRNA Diagnostics.
Advanced Materials ( IF 27.4 ) Pub Date : 2019-10-30 , DOI: 10.1002/adma.201905311
Richard Bruch 1, 2 , Julia Baaske 3 , Claire Chatelle 3 , Mailin Meirich 1 , Sibylle Madlener 4 , Wilfried Weber 3 , Can Dincer 1, 2, 5 , Gerald Anton Urban 1, 6
Affiliation  

Noncoding small RNAs, such as microRNAs, are becoming the biomarkers of choice for multiple diseases in clinical diagnostics. A dysregulation of these microRNAs can be associated with many different diseases, such as cancer, dementia, and cardiovascular conditions. The key for effective treatment is an accurate initial diagnosis at an early stage, improving the patient's survival chances. In this work, the first clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a-powered microfluidic, integrated electrochemical biosensor for the on-site detection of microRNAs is introduced. Through this unique combination, the quantification of the potential tumor markers microRNA miR-19b and miR-20a is realized without any nucleic acid amplification. With a readout time of 9 min and an overall process time of less than 4 h, a limit of detection of 10 pm is achieved, using a measuring volume of less than 0.6 µL. Furthermore, the feasibility of the biosensor platform to detect miR-19b in serum samples of children, suffering from brain cancer, is demonstrated. The validation of the obtained results with a standard quantitative real-time polymerase chain reaction method shows the ability of the electrochemical CRISPR-powered system to be a low-cost, easily scalable, and target amplification-free tool for nucleic acid based diagnostics.

中文翻译:

CRISPR / Cas13a供电的电化学微流生物传感器,用于无核酸扩增的miRNA诊断。

非编码小RNA(例如microRNA)正成为临床诊断中多种疾病选择的生物标记。这些microRNA的失调可能与许多不同的疾病有关,例如癌症,痴呆症和心血管疾病。有效治疗的关键是在早期进行准确的初始诊断,从而提高患者的生存机会。在这项工作中,介绍了第一个成簇的规则间隔的短回文重复序列(CRISPR)/ Cas13a驱动的微流集成电化学生物传感器,用于现场检测microRNA。通过这种独特的组合,无需任何核酸扩增即可实现潜在肿瘤标记物microRNA miR-19b和miR-20a的定量。读取时间为9分钟,总处理时间少于4小时,使用小于0.6 µL的测量体积可达到10 pm的检测极限。此外,证明了生物传感器平台检测患有脑癌的儿童血清样品中的miR-19b的可行性。用标准的实时定量聚合酶链反应方法对获得的结果进行验证,证明了基于CRISPR的电化学系统具有低成本,易于扩展和无目标扩增的免费功能,可用于基于核酸的诊断。
更新日期:2019-12-18
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