Pharmaceutical Engineering Research Center, College of Pharmacy, Chongqing Medical University, Chongqing, China; Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Institute for Viral Hepatitis, Department of Infectious Diseases, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.
Department of Respiratory Disease, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Biomaterials. 2017 Jul;133:11-19. doi: 10.1016/j.biomaterials.2017.04.010. Epub 2017 Apr 12.
Tuberculosis caused by Mycobacterium tuberculosis (MTB) is still a major threat to global public health. However, the existing methods for MTB detection are usually complicated and time consuming with unsatisfactory sensitivity and specificity. In this work, a relatively simple and ultrasensitive electrochemical aptasensor based on novel signal generation and amplification was constructed for the determination of MTB antigen MPT64. The coil-like fullerene (C)-doped polyaniline (C-PAn) nanohybrids with large surface area, abundant active groups and excellent electric performance were synthesized and used both as new redox nanoprobe and catalyst for the generation and amplification of electrochemical signal for the first time. Then gold nanoparticles decorated C-PAn nanocomposites (GNPs-C-PAn) were labeled with signal aptamer to form the tracer label. After the sandwich reaction of target MPT64 antigen between capture aptamer and the tracer label, a distinguishing detection signal of C-PAn would be observed. Moreover, the detection signal could be enormously enhanced towards the efficient electrocatalytic oxidation of ascorbic acid based on C-PAn, resulting in further improvement of the sensitivity. With the excellent redox and electrocatalytic activity of C-PAn, a wide detection linear range from 0.02 to 1000 pg/mL was obtained with a detection limit of 20 fg/mL for MPT64. The proposed aptasensor showed high selectivity to target antigen compared with possible interfering substances. More importantly, it also exhibited excellent specificity and sensitivity for MPT64 detection in serum samples of tuberculosis patients, which provided a rapid and efficient detection method for MTB infection.
结核分枝杆菌(MTB)引起的结核病仍然是全球公共卫生的主要威胁。然而,现有的 MTB 检测方法通常比较复杂,耗时较长,并且灵敏度和特异性不理想。在这项工作中,构建了一种相对简单且超灵敏的基于新型信号产生和放大的电化学适体传感器,用于检测结核分枝杆菌抗原 MPT64。具有大表面积、丰富的活性基团和优异的电性能的管状富勒烯(C)掺杂聚苯胺(C-PAn)纳米杂化物被首次用作新的氧化还原纳米探针和催化剂,用于电化学信号的产生和放大。然后,金纳米粒子修饰的 C-PAn 纳米复合材料(GNPs-C-PAn)被信号适体标记,形成示踪标记。在目标 MPT64 抗原与捕获适体和示踪标记之间发生三明治反应后,将观察到 C-PAn 的明显检测信号。此外,由于基于 C-PAn 的抗坏血酸的高效电催化氧化,检测信号可以大大增强,从而进一步提高了灵敏度。由于 C-PAn 的出色的氧化还原和电催化活性,该适体传感器获得了从 0.02 到 1000 pg/mL 的宽检测线性范围,检测限为 20 fg/mL。与可能的干扰物质相比,该适体传感器对目标抗原表现出高选择性。更重要的是,它还在结核患者的血清样本中对 MPT64 检测表现出优异的特异性和灵敏度,为 MTB 感染提供了一种快速有效的检测方法。