基于环介导等温扩增技术水产品中副溶血弧菌快速检测方法的建立与应用

Rapid detection of Vibrio parahaemolyticus in aquatic products using a loop-mediated isothermal amplification method

本试验基于环介导等温扩增(LAMP)技术,以副溶血弧菌toxR基因为靶基因,设计两对LAMP引物F3/B3和FIP/BIP,建立了副溶血弧菌LAMP检测方法,60min内即可完成检测。该LAMP方法特异性强、灵敏度高,对纯培养副溶血弧菌的检测灵敏度为42CFU/mL,对污染食品中副溶血弧菌的检测灵敏度为67CFU/mL。实践应用证明,本试验建立的副溶血弧菌LAMP检测方法操作简单、检测结果准确,具有良好的实用性。

In this work, loop-mediated isothermal amplification （LAMP） technique was introduced to develop the method for rapid detection of V. parahaemolyticus. Two pairs of primers F3/B3 and FIP/BIP were designed according to the toxR gene of V. parahaernolyticus, and then the LAMP method for V. parahaemolyticus was developed. The detection for V. parahaernolyticus could be completed in 60min by the method. The LAMP method was highly specific to V. parahaemolyticus with a sensitivity of 42 CFU/ mL for cultivated V. parahaernolyticus and 67CFU/mL for its contaminated food. Practical applications showed that the LAMP method developed in this work was a sensitive, rapid and simple tool for the detec-tion of V. parahaemolyticus.