基于生物发光的高灵敏度三磷酸腺苷检测研究

Highly-Sensitive Detecting of Adenosine Triphosphate Based on Bioluminescence

在微生物检测和食品安全控制中需要对低浓度的三磷酸腺苷(ATP)做出快速准确的测定,基于ATP光学反应原理,设计了一种微量光学反应池,结合萤火虫素-萤火虫素酶发光技术,构建了一种用于现场检测的高灵敏度ATP光学传感器系统。传感器响应波长为550nm,ATP生物发光反应适宜pH为7.8,在优化实验条件下,发光强度和ATP浓度在对数坐标下呈线性相关,检出限可达1×10-16mol/LATP,检测样品仅需30μL,测量时间60s。这种检测方法具有响应范围宽,测试简便,快速可靠等特点,适用于ATP现场快速检测,具有很好的应用前景。

In microorganism determination and food safety control, the low concentration of adenosine triphosphate （ATP） need be detected rapidly and accurately because of the special requirements and applications. Based on the principle of ATP bioluminescence reaction, a micro-optical-sensing cell was designed and a kind of highly-sensitive optical sensor system for field detecting of ATP has been constructed with luciferin-luciferase hioluminescence technique. The response wavelength of the sensor was 550 nm and the appropriate pH for the ATP bioluminescence reaction was 7.8. Under the optimized experimental conditions, the results indicated that a favorable linearity between the ATP concentration and the luminescence intensity was presented in loglog relationship, 10-16 mol/L ATP could be detected. The sampling volume was 30 uL and the whole testing time was 60 s. The test results revealed that the range, simple operation and fast detection, which method possessed many advantages, including wide linear have good prospect in the practical application on-site.