State Key Laboratory of Pharmaceutical Biotechnology and Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing 210093, PR China.
Department of Oncology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, PR China.
Talanta. 2018 Feb 1;178:89-93. doi: 10.1016/j.talanta.2017.09.011. Epub 2017 Sep 5.
At present, the accuracy of clinical hepatocellular carcinoma (HCC) diagnosis needs to be further improved. In this work, two kinds of serological biomarker species, microRNA and protein biomarker, have been detected simultaneously to identify HCC. Herein, a dual-aptamer hairpin DNA oligonucleotide is designed as the electrochemical sensing probe (ESP) to achieve this goal. The hairpin-structured DNA probe consists microRNA-16 (miR-16) complementary sequence and alpha fetoprotein (AFP) aptamer sequence, so it can both capture miR-16 and AFP. Once it hybridizes with miR-16, the hairpin structure is unlocked so that the terminal modified signal molecule (methylene blue, MB) would give a decreased electrochemical signal. Meanwhile, once it recognizes AFP, concanavalin A (ConA) modified silver nanoparticles (AgNPs) can bind to AFP at the sensing surface. An obvious electrochemical signal of AgNPs can thus be generated for AFP detection. In this way, one-step and simultaneous detection of miRNA-16 and AFP can easily be realized by collecting the two sensitive and non-interfering electrochemical signals. Compared with traditional single biomarker detection methods, this assay strategy can improve the accuracy of HCC by monitoring two kinds of serological biomarkers species. Besides, this novel electrochemical biosensor based on ESP is simple, low-cost and efficient, which make it promising to improve the accuracy and specificity for the diagnosis HCC in the future.
目前，临床肝细胞癌 (HCC) 的诊断准确性需要进一步提高。在这项工作中，同时检测了两种血清生物标志物，即 microRNA 和蛋白质生物标志物，以识别 HCC。在这里，设计了一种双适体发夹 DNA 寡核苷酸作为电化学传感探针 (ESP) 来实现这一目标。发夹结构的 DNA 探针由 microRNA-16 (miR-16) 互补序列和甲胎蛋白 (AFP) 适体序列组成，因此它既能捕获 miR-16 又能捕获 AFP。一旦与 miR-16 杂交，发夹结构就会被解锁，从而使末端修饰的信号分子 (亚甲基蓝，MB) 的电化学信号减弱。同时，一旦它识别 AFP，修饰后的 ConA 的银纳米粒子 (AgNPs) 就可以与传感表面上的 AFP 结合。因此，AgNPs 的明显电化学信号可以用于 AFP 的检测。这样，通过收集两种敏感且互不干扰的电化学信号，就可以轻松实现 miRNA-16 和 AFP 的一步和同时检测。与传统的单一生物标志物检测方法相比，该检测策略通过监测两种血清生物标志物种类，可提高 HCC 的诊断准确性。此外，这种基于 ESP 的新型电化学生物传感器简单、低成本且高效，有望在未来提高 HCC 的诊断准确性和特异性。
