College of Life Science, Hebei Normal University, Shijiazhuang, 050024, PR China; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, PR China.
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, PR China.
Anal Chim Acta. 2024 Mar 22;1295:342306. doi: 10.1016/j.aca.2024.342306. Epub 2024 Jan 30.
Point-of-care quantitative analysis of tracing microRNA disease-biomarkers remains a great challenge in the clinical diagnosis. In this paper, we developed a portable fluorescent lateral flow assay for ultrasensitive quantified detection of acute myocardial infarction related microRNAs in bio-samples. SiO@DQD (bilayer quantum dots assembly with SiO core) based fluorescent lateral flow strip was fabricated as the analysis tool. In order to quantify the tracing microRNA in biosamples, a catalytic hairpin assembly and CRISPR/Cas12a cascade amplification method was performed and combined with the fabricated SiO@DQD lateral flow strip. Thus, our platform gathered double advantages of portability and ultrasensitive quantification. Based on our strips, target myocardial biomarker microRNA-133a can be detected with a detection limit of 0.32 fM, which was almost 1000-fold sensitive compared with previous reported microRNAs-lateral flow strips. Significantly, this portable fluorescent strip can directly detect microRNAs in serum without any pretreatment and PCR amplification steps. When spiked in serum samples, a recovery of 99.65 %-102.38 % can be obtained. Therefore, our method offers a potential tool for ultrasensitive quantification of diseases related microRNA in the point-of-care diseases diagnosis field.
即时检测痕量microRNA 疾病生物标志物仍然是临床诊断中的一大挑战。本文中，我们开发了一种便携式荧光侧向流动分析方法，用于超灵敏定量检测生物样本中的急性心肌梗死相关 microRNA。以基于 SiO@DQD（具有 SiO 核的双层量子点组装）的荧光侧向流动条作为分析工具。为了定量检测生物样本中的追踪 microRNA，我们进行了催化发夹组装和 CRISPR/Cas12a 级联扩增方法，并与所制备的 SiO@DQD 侧向流动条相结合。因此，我们的平台汇集了便携性和超灵敏定量的双重优势。基于我们的条带，可以检测到靶心肌生物标志物 microRNA-133a，检测限为 0.32 fM，与以前报道的 microRNAs-侧向流动条相比，灵敏度提高了近 1000 倍。值得注意的是，这种便携式荧光条可以直接在未经任何预处理和 PCR 扩增步骤的血清中检测 microRNAs。当在血清样品中加入时，可以获得 99.65%-102.38%的回收率。因此，我们的方法为即时检测疾病相关 microRNA 的超灵敏定量提供了一种潜在的工具，可用于疾病诊断领域。
