Chen Meiyan, Kang Qiongdan, Zhang Annan, Lin Shanti, Chen Zhangxing
Department of Gastroenterology, Xiamen University Affiliated Chenggong Hospital, Xiamen City, Fujian Province 361003, China.
ACS Omega. 2024 Dec 17;9(52):51157-51162. doi: 10.1021/acsomega.4c06859. eCollection 2024 Dec 31.
MicroRNAs (miRNAs), which play critical roles in regulating gene expression and cell functions, are recognized as potential biomarkers for various human diseases, including gastric ulcers. The reliable, specific, and sensitive detection of miRNA is highly recommended for the clinical diagnosis and therapy of different diseases. Herein, we depict a label-free and low-background fluorescent assay for the highly sensitive detection of miRNAs by coupling target miRNA-triggered cyclization of a padlock, circular padlock-mediated catalytic hairpin assembly (CHA), and primer exchange reaction (PER)-assisted signal generation. The padlock probe recognizes the target miRNA, forming a circular padlock that subsequently facilitates the CHA. The subsequent PER process generates substantial quantities of G-quadruplex sequences that rapidly combine with thioflavin T to create substantial fluorescence, thereby enabling the highly sensitive detection of the target miRNA. This method demonstrated significant potential for the early diagnosis of diseases such as gastric ulcers, as it could conclude the detection process in human serum samples within hours.
微小RNA（miRNA）在调节基因表达和细胞功能方面发挥着关键作用，被认为是包括胃溃疡在内的各种人类疾病的潜在生物标志物。强烈建议对miRNA进行可靠、特异且灵敏的检测，以用于不同疾病的临床诊断和治疗。在此，我们描述了一种无标记且背景低的荧光检测方法，用于通过耦合靶标miRNA触发的锁式探针环化、环状锁式探针介导的催化发夹组装（CHA）和引物交换反应（PER）辅助信号产生来高度灵敏地检测miRNA。锁式探针识别靶标miRNA，形成环状锁式探针，随后促进CHA。随后的PER过程产生大量的G-四链体序列，这些序列迅速与硫黄素T结合产生大量荧光，从而能够高度灵敏地检测靶标miRNA。该方法在胃溃疡等疾病的早期诊断中显示出巨大潜力，因为它可以在数小时内完成人血清样本中的检测过程。
