Borthwick Kathryn A J, Love Tracey E, McDonnell Martin B, Coakley W Terence
Cardiff School of Biosciences, Cardiff University, Wales, UK.
Anal Chem. 2005 Nov 15;77(22):7242-5. doi: 10.1021/ac050576c.
Ultrasonic cavitation was employed to enhance sensitivity of bacterial spore immunoassay detection, specifically, enzyme-linked immunosorbent assay (ELISA) and resonant mirror (RM) sensing. Bacillus spore suspensions were exposed to high-power ultrasound in a tubular sonicator operated at 267 kHz in both batch and flow modes. The sonicator was designed to deliver high output power and is in a form that can be cooled efficiently to avoid thermal denaturation of antigen. The 30-s batch and cooled flow (0.3 mL/min) sonication achieved an approximately 20-fold increase in ELISA sensitivity compared to unsonicated spores by ELISA. RM sensing of sonicated spores achieved detection sensitivity of approximately 10(6) spores/mL, whereas unsonicated spores were undetectable at the highest concentration tested. Improvements in detection were associated with antigen released from the spores. Equilibrium temperature increase in the tubular sonicator was limited to 14 K after 30 min and was maintained for 6 h with cooling and flow (0.3 mL/min). The work described here demonstrates the utility of the tubular sonicator for the improvement in the sensitivity of the detection of spores and its suitability as an in-line component of a rapid detection system.
采用超声空化技术提高细菌芽孢免疫分析检测的灵敏度，具体而言，是提高酶联免疫吸附测定（ELISA）和共振镜（RM）传感的灵敏度。将芽孢杆菌芽孢悬液在以267 kHz运行的管式超声处理器中以分批和流动模式暴露于高功率超声下。该超声处理器设计用于提供高输出功率，其形式能够有效冷却以避免抗原热变性。与未超声处理的芽孢相比，30秒的分批超声处理和冷却流动（0.3 mL/分钟）超声处理使ELISA检测灵敏度提高了约20倍。对超声处理后的芽孢进行RM传感，检测灵敏度达到约10(6)个芽孢/毫升，而在测试的最高浓度下，未超声处理的芽孢无法检测到。检测的改善与芽孢释放的抗原有关。管式超声处理器中的平衡温度在30分钟后升高限制在14 K，并通过冷却和流动（0.3 mL/分钟）保持6小时。此处描述的工作证明了管式超声处理器在提高芽孢检测灵敏度方面的实用性及其作为快速检测系统在线组件的适用性。
