Department of Advanced Sciences and Technology for Biomedical Sensors, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549, Japan; E-Mail:
Graduate School of Bionics, Tokyo University of Technology, 1404-1, Katakura-machi, Hachioji-shi, Tokyo 192-0982, Japan; E-Mails:
Biosensors (Basel). 2013 Mar 8;3(1):120-31. doi: 10.3390/bios3010120. eCollection 2013 Mar.
Dynamic fluoroimmunoassay with a flow-through system using optical fiber probes consisting of polystyrene was developed and applied to a quantitative detection of E. coli O157:H7. The system measures E. coli as fluorescence of sandwich-type immune complexes formed by capture antibodies immobilized on the surface of the probe, E. coli cells, and fluorescently labeled detection antibodies. Excitation was carried out using an evanescent wave from the probe. Resulting fluorescence recoupled into the probe was detected by a photodiode. The assay system was constructed with a flow cell which was available for sequential injection of experimental reagents. In vitro characterization was performed using the flow cell, and the calibration range of E. coli O157:H7 was from 10(3) to 10(7) cells/mL. The measurement for each sample was completed within 12 min. Furthermore, it was also possible to estimate the concentrations of E. coli O157:H7 by the increasing rate of fluorescence during binding reaction of detection antibodies to antigens. This minimized the time for measurement down to 6 min. The system is suitable for rapid and direct determination for microorganisms or bacteria in food, clinical, and environmental sources.
采用光纤探针的流动式动态荧光免疫分析系统被开发出来并应用于定量检测大肠杆菌 O157:H7。该系统通过测量由固定在探针表面的捕获抗体、大肠杆菌细胞和荧光标记的检测抗体形成的三明治型免疫复合物的荧光来检测大肠杆菌。激发是通过探针的消逝波进行的。通过光电二极管检测重新耦合到探针中的荧光。该检测系统由一个流动池构成，可用于连续注入实验试剂。在体外使用流动池进行了特征描述，大肠杆菌 O157:H7 的校准范围为 10(3)到 10(7)个细胞/mL。每个样本的测量在 12 分钟内完成。此外，还可以通过检测抗体与抗原结合反应过程中荧光的增加率来估计大肠杆菌 O157:H7 的浓度。这将测量时间缩短至 6 分钟。该系统适用于食品、临床和环境来源中微生物或细菌的快速直接检测。
