Yuan Kaisong, Mei Qingsong, Guo Xinjie, Xu Youwei, Yang Danting, Sánchez Beatriz Jurado, Sheng Bingbing, Liu Chusheng, Hu Ziwei, Yu Guangchao, Ma Hongming, Gao Hao, Haisch Christoph, Niessner Reinhard, Jiang Zhengjin, Jiang Zhengjing, Zhou Haibo
Institute of Pharmaceutical Analysis , College of Pharmacy , Jinan University , Guangzhou , Guangdong 510632 , China . Email:
Department of Analytical Chemistry , Physical Chemistry and Chemical Engineering , University of Alcala , Alcala de Henares E-28871 , Madrid , Spain.
Chem Sci. 2018 Nov 2;9(47):8781-8795. doi: 10.1039/c8sc04637a. eCollection 2018 Dec 21.
In this study, a new biosensor based on a sandwich structure has been developed for the isolation and detection of multiple bacterial pathogens magnetic separation and SERS tags. This novel assay relies on antimicrobial peptide (AMP) functionalized magnetic nanoparticles as "capturing" probes for bacteria isolation and gold coated silver decorated graphene oxide (Au@Ag-GO) nanocomposites modified with 4-mercaptophenylboronic acid (4-MPBA) as SERS tags. When different kinds of bacterial pathogens are combined with the SERS tags, the "fingerprints" of 4-MPBA show corresponding changes due to the recognition interaction between 4-MPBA and different kinds of bacterial cell wall. Compared with the label-free SERS detection of bacteria, 4-MPBA here can be used as an internal standard (IS) to correct the SERS intensities with high reproducibility, as well as a Raman signal reporter to enhance the sensitivity and amplify the differences among the bacterial "fingerprints". Thus, three bacterial pathogens (, and ) were successfully isolated and detected, with the lowest concentration for each of the strains detected at just 10 colony forming units per mL (CFU mL). According to the changes in the "fingerprints" of 4-MPBA, three bacterial strains were successfully discriminated using discriminant analysis (DA). In addition, the AMP modified FeONPs feature high antibacterial activities, and can act as antibacterial agents with low cellular toxicology in the long-term storage of blood for future safe blood transfusion applications. More importantly, this novel method can be applied in the detection of bacteria from clinical patients who are infected with bacteria. In the validation analysis, 97.3% of the real blood samples (39 patients) could be classified effectively (only one patient infected with was misclassified). The multifunctional biosensor presented here allows for the simultaneous isolation, discrimination and killing of bacteria, suggesting its high potential for clinical diagnosis and safe blood transfusions.
在本研究中，基于三明治结构开发了一种新型生物传感器，用于通过磁分离和表面增强拉曼散射（SERS）标签分离和检测多种细菌病原体。这种新型检测方法依赖于抗菌肽（AMP）功能化的磁性纳米颗粒作为细菌分离的“捕获”探针，以及用4-巯基苯硼酸（4-MPBA）修饰的金包覆银修饰的氧化石墨烯（Au@Ag-GO）纳米复合材料作为SERS标签。当不同种类的细菌病原体与SERS标签结合时，由于4-MPBA与不同种类细菌细胞壁之间的识别相互作用，4-MPBA的“指纹”会出现相应变化。与无标记的细菌SERS检测相比，此处的4-MPBA可用作内标（IS）以高度可重复地校正SERS强度，同时作为拉曼信号报告分子以提高灵敏度并放大细菌“指纹”之间的差异。因此，成功分离并检测到三种细菌病原体（、和），每种菌株检测到的最低浓度仅为每毫升10个菌落形成单位（CFU/mL）。根据4-MPBA“指纹”的变化，使用判别分析（DA）成功区分了三种细菌菌株。此外，AMP修饰的FeONPs具有高抗菌活性，并且在血液的长期储存中可作为细胞毒理学低的抗菌剂，用于未来的安全输血应用。更重要的是，这种新方法可应用于检测感染细菌的临床患者的细菌。在验证分析中，97.3%的真实血液样本（39名患者）能够有效分类（仅一名感染的患者被错误分类）。此处展示的多功能生物传感器能够同时分离、区分和杀灭细菌，表明其在临床诊断和安全输血方面具有很高的潜力。
