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基于发夹变构分子信标的级联扩增用于有效检测肺癌相关微小RNA

Hairpin allosteric molecular beacons-based cascaded amplification for effective detection of lung cancer-associated microRNA.

作者信息

Zheng Cheng, Hu Xuemei, Sun Shujuan, Zhu Lingye, Wang Ning, Zhang Jing, Huang Guoqiao, Wang Yuzhe, Huang Xiaoying, Wang Liangxing, Shen Zhifa

机构信息

Division of Pulmonary Medicine, The First Affiliated Hospital of Wenzhou Medical University, Key Laboratory of Heart and Lung, Wenzhou Medical University, Wenzhou, Zhejiang, 325000, China; Zhejiang Provincial Key Laboratory of Medical Genetics, Key Laboratory of Laboratory Medicine, Ministry of Education, China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, 325035, China.

Zhejiang Provincial Key Laboratory of Medical Genetics, Key Laboratory of Laboratory Medicine, Ministry of Education, China, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, 325035, China.

出版信息

Talanta. 2022 Jul 1;244:123412. doi: 10.1016/j.talanta.2022.123412. Epub 2022 Mar 28.

Abstract

Lung cancer with worldwide distribution, high incidence and low survival rate is significantly and increasingly threatening human health. Thus, the specific detection of lung cancer-associated biomarkers is of crucial importance in early diagnosis and treatment. In this work, taking microRNA-21 as an example, a biosensor is proposed via a stimuli-induced strand displacement amplification (SDA) and cascade signal amplification with the assistance of polymerase. An allosteric molecular beacon (MB) with chemical modification is designed to emit the enhanced fluorescent signal in presence of microRNA target. The sensing system possesses a linear calibration curve from 5 pM to 40 nM with the limit of detection (LOD) of 0.7 pM and displays good specificity to discriminate coexisting microRNAs. In addition, the feasibility is confirmed by performing the detection of miRNA-21 extracted from non-small cell lung cancer (NSCLC) A549 cells, and a good recovery is achieved in complex human serum sample. Therefore, the miRNA-triggered cascaded amplification would be crucial strategy to facilitate microRNA analysis in the biological detection and broad clinical applications.

摘要

肺癌在全球范围内分布广泛,发病率高且生存率低,对人类健康构成了日益严重的威胁。因此,肺癌相关生物标志物的特异性检测对于早期诊断和治疗至关重要。在这项工作中,以微小RNA-21为例,通过刺激诱导链置换扩增(SDA)和在聚合酶辅助下的级联信号放大,提出了一种生物传感器。设计了一种具有化学修饰的变构分子信标(MB),以在存在微小RNA靶标的情况下发出增强的荧光信号。该传感系统具有从5 pM到40 nM的线性校准曲线,检测限(LOD)为0.7 pM,并且对区分共存的微小RNA显示出良好的特异性。此外,通过对从非小细胞肺癌(NSCLC)A549细胞中提取的miRNA-21进行检测,证实了该方法的可行性,并且在复杂的人血清样本中实现了良好的回收率。因此,miRNA触发的级联扩增将是促进生物检测和广泛临床应用中微小RNA分析的关键策略。

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