Key Laboratory of Laboratory Medicine, Ministry of Education of China, and Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China.
Key Laboratory of Laboratory Medicine, Ministry of Education of China, and Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China; Cancer Metastasis Alert and Prevention Center, Biopharmaceutical Photocatalysis of State Key Laboratory of Photocatalysis on Energy and Environment, and Fujian Provincial Key Laboratory of Cancer Metastasis Chemoprevention and Chemotherapy, Fuzhou University, Fuzhou 350002, China.
Talanta. 2018 Sep 1;187:365-369. doi: 10.1016/j.talanta.2018.05.035. Epub 2018 May 10.
The development of rapid approaches to detect prognostic markers is significant in reducing the morbidity and mortality of cancer. In this paper, we describe a rapid and specific biosensing platform for target DNA (p53 gene as a model) detection based on reverse strand displacement amplification (R-SDA). When the p53 gene is added, multifuctional molecular beacon (MMB) is unfolded via the hybridization with p53 gene. With the assist of Klenow fragment (KF) and Nt.BbvCI (the nicking endonuclease), p53 gene recycling could be initiated and considerable amount of complementary sequences for the MMBs (Nicked fragments, NFs) could be formed, generating enhanced fluorescence signal. Using this amplification strategy, the proposed biosensor displays the detection limit of 1 nM and a wide linear range from 1 to 100 nM, even if only one type of probe is involved. Notably, remarkable detection specificity for single-base mismatched target p53 gene is achieved. Moreover, the described biosensor also exhibited the stability in real biological samples (human serum). The rapid detection strategy can be performed less than 30 min without harsh reaction conditions or expensive nanoparticles. This biosensor shows great potential for application in clinic assay, especially, for early cancer diagnosis.
快速检测预后标志物对于降低癌症的发病率和死亡率具有重要意义。本文描述了一种基于反向链置换扩增（R-SDA）的用于靶 DNA（以 p53 基因为模型）检测的快速和特异的生物传感平台。当加入 p53 基因时，多功能分子信标（MMB）通过与 p53 基因杂交而展开。在 Klenow 片段（KF）和 Nt.BbvCI（切口内切酶）的辅助下，可以启动 p53 基因的循环，并且可以形成大量用于 MMB 的互补序列（缺口片段，NFs），从而产生增强的荧光信号。使用这种扩增策略，所提出的生物传感器显示出 1 nM 的检测限和 1 至 100 nM 的宽线性范围，即使只涉及一种探针也是如此。值得注意的是，该生物传感器实现了对单碱基错配靶 p53 基因的显著检测特异性。此外，所描述的生物传感器在真实生物样品（人血清）中也表现出稳定性。快速检测策略可以在不到 30 分钟的时间内完成，而无需苛刻的反应条件或昂贵的纳米颗粒。该生物传感器在临床检测中具有很大的应用潜力，特别是在早期癌症诊断方面。
