Ondera Thomas J, Hamme Ashton T
Department of Chemistry and Biochemistry, Jackson State University, 1400 J R Lynch street, Jackson, MS 39217, USA.
Analyst. 2015 Dec 7;140(23):7902-11. doi: 10.1039/c5an00497g.
A rapid, sensitive and quantitative immunoassay for the targeted detection and decontamination of E. coli based on Fe3O4 magnetic nanoparticles (MNPs) and plasmonic popcorn-shaped gold nanostructure attached single-walled carbon nanotubes (AuNP@SWCNT) is presented. The MNPs were synthesized as the support for a monoclonal antibody (mAb@MNP). E. coli (49979) was captured and rapidly preconcentrated from the sample with the mAb@MNP, followed by binding with Raman-tagged concanavalin A-AuNP@SWCNTs (Con A-AuNP@SWCNTs) as detector nanoprobes. A Raman tag 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) generated a Raman signal upon 670 nm laser excitation enabling the detection and quantification of E. coli concentration with a limit of detection of 10(2) CFU mL(-1) and a linear logarithmic response range of 1.0 × 10(2) to 1.0 × 10(7) CFU mL(-1). The mAb@MNP could remove more than 98% of E. coli (initial concentration of 1.3 × 10(4) CFU mL(-1)) from water. The potential of the immunoassay to detect E. coli bacteria in real water samples was investigated and the results were compared with the experimental results from the classical count method. There was no statistically significant difference between the two methods (p > 0.05). Furthermore, the MNP/AuNP@SWCNT hybrid system exhibits an enhanced photothermal killing effect. The sandwich-like immunoassay possesses potential for rapid bioanalysis and the simultaneous biosensing of multiple pathogenic agents.
本文介绍了一种基于Fe3O4磁性纳米颗粒(MNPs)和附着有等离子体爆米花状金纳米结构的单壁碳纳米管(AuNP@SWCNT)的快速、灵敏且定量的免疫分析方法,用于靶向检测和净化大肠杆菌。合成了MNPs作为单克隆抗体的载体(mAb@MNP)。用mAb@MNP从样品中捕获并快速预富集大肠杆菌(49979),随后与作为检测纳米探针的拉曼标记伴刀豆球蛋白A-AuNP@SWCNTs(Con A-AuNP@SWCNTs)结合。拉曼标签5,5'-二硫代双(2-硝基苯甲酸)(DTNB)在670 nm激光激发下产生拉曼信号,能够检测和定量大肠杆菌浓度,检测限为10(2) CFU mL(-1),线性对数响应范围为1.0×10(2)至1.0×10(7) CFU mL(-1)。mAb@MNP可以从水中去除超过98%的大肠杆菌(初始浓度为1.3×10(4) CFU mL(-1))。研究了该免疫分析方法检测实际水样中大肠杆菌的潜力,并将结果与经典计数法的实验结果进行了比较。两种方法之间没有统计学上的显著差异(p > 0.05)。此外,MNP/AuNP@SWCNT混合系统表现出增强的光热杀伤效果。这种三明治式免疫分析方法具有快速生物分析和同时检测多种病原体的潜力。