副溶血弧菌现场检测系统的研发及应用

Development and Application of Point-of-Care Test System for Detection of Vibrio parahaemolyticus

为满足现场检测海产品中的副溶血弧菌(Vibrio parahaemolyticus)的需求,本文建立了一种基于电化学检测和环介导等温扩增方法的副溶血弧菌现场检测系统,设备核心电路的尺寸仅为7 cm×4 cm,质量为30 g,易于携带,可以在30 min内出检测结果,且可以对样本中的副溶血弧菌进行定量分析,检测限可达103 CFU/mL。同时,利用该系统对加标虾样品中副溶血弧菌进行检测,检测限低至3.6×103 CFU/g。随着核酸扩增反应的进行,核酸扩增体系的pH降低,本文制备的pH敏感电极可以实时监测核酸扩增体系的pH微量变化,精度为0.016个pH单位,本系统的检测限与荧光检测的检测限一致。本系统具有便携、快速、特异性好、灵敏度高、成本低、可现场检测等优点,本研究有利于食品安全中分子诊断技术的基层下沉,满足公共卫生安全的战略需求。

In order to meet the requirements of the on-site detection of Vibrio parahaemolyticus in seafood,an point-of-care test system for V.parahaemolyticus based on electrochemical detection and loop-mediated isothermal amplification was established in this study.The core circuit of the device is only 7 cm×4 cm in size and weighs 30 g.The system is easy to carry.The detection result can be obtained within 30 min.The detection limit of V.parahaemolyticus in samples can reach 103 CFU/mL.At the same time,the system is used to detect V.parahaemolyticus in the stained shrimp samples,with a detection limit as low as 3.6×103 CFU/g.With the progress of the nucleic acid amplification reaction,the pH of the nucleic acid amplification system decreased.The pH sensitive electrode prepared in this study can monitor the pH change of the nucleic acid amplification system in real time with an accuracy of 0.016 pH units.Therefore,the detection limit of this system can be consistent with that of fluorescence detection.The system has the advantages of portability,rapidness,high sensitivity,low cost,on-site detection among others,which is conducive to sink downward to the basic level of molecular diagnosis technology in food safety,and meets the important national strategic needs of public health safety.