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 400016, China.
Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
Biosens Bioelectron. 2019 Feb 15;127:167-173. doi: 10.1016/j.bios.2018.12.012. Epub 2018 Dec 13.
Herein, a novel and pragmatic electrochemiluminescence (ECL) biosensing method was developed for ultrasensitive and specific detection of Group B Streptococci (GBS) by combining self-enhanced luminol complex functionalized CuMn-CeO (CuMn-CeO-PEI-luminol) with MNAzyme-mediated target-recycling amplification. First, the efficient self-enhanced PEI-luminol luminophore was prepared by combining PEI co-reactant with luminol in one molecular, which shortened electron transfer distance and enhanced ECL signal. And CuMn-CeO was applied to load a large number of PEI-luminol and strengthen luminous efficiency of luminol by the high catalytic activity toward HO oxidation. Then, target-driven MNAzyme system was used to realize the circulation of GBS nucleic acid sequence, producing plentiful triggers to initiate the hybridization reaction on the surface of electrode. The developed enzyme-free ECL biosensor showed ultra-sensitivity for target DNA detection with detection limits of 68 aM (synthetic DNA) and 5 × 10 CFU mL (genomic DNA extracted from GBS strain). More importantly, this biosensor was successfully applied for detection of genomic DNA of GBS extracted from clinical vaginal/anal swabs as low as 320 copies. Thus, this proposed strategy might be an pragmatic ECL platform for ultrasensitive and specific detection of GBS in clinical vaginal/anal swabs.
在此,通过将自增强的 PEI-鲁米诺发光体与 MNAzyme 介导的靶标循环扩增相结合,开发了一种新颖且实用的用于超灵敏和特异性检测 B 族链球菌(GBS)的电化学发光(ECL)生物传感方法。首先,通过将 PEI 共反应物与鲁米诺在一个分子中结合,制备了高效的自增强 PEI-鲁米诺发光体,缩短了电子转移距离并增强了 ECL 信号。并且,CuMn-CeO 被应用于负载大量的 PEI-鲁米诺,并通过对 HO 氧化的高催化活性增强鲁米诺的发光效率。然后,目标驱动的 MNAzyme 系统用于实现 GBS 核酸序列的循环,产生大量的触发物来启动电极表面的杂交反应。所开发的无酶 ECL 生物传感器对目标 DNA 检测具有超灵敏性,检测限分别为 68 aM(合成 DNA)和 5 × 10 CFU mL(从 GBS 菌株提取的基因组 DNA)。更重要的是,该生物传感器成功地应用于检测临床阴道/肛门拭子中提取的 GBS 基因组 DNA,其检测下限低至 320 拷贝。因此,该策略可能是一种用于临床阴道/肛门拭子中 GBS 超灵敏和特异性检测的实用 ECL 平台。