提升副溶血性弧菌检验前增菌速度

Increase the Rate of Pre-enrichment Before Vibrio parahaemolyticus Tested

为有效缩短前增菌时间并打破副溶血性弧菌快检的技术瓶颈,研究通过优化培养基配方和培养条件,探索提升副溶血性弧菌前增菌速度的技术方法。通过单因素和响应面分析确定最适培养基配方为:大豆蛋白胨7.4 g/L、酵母浸粉9.8 g/L、牛心浸粉20.6 g/L,Na Cl 30.0 g/L,p H8.0。最佳培养条件为:温度36℃,转速180 r/min。应用优化的增菌培养方法,对副溶血性弧菌增菌培养6h后,即可获得满足荧光聚合酶链式反应(Polymerase chain reaction,PCR)检测灵敏度要求的增菌液。在添加食品基质的样本中,与GB 4789.7增菌方法相比,优化的增菌培养方法表现出了较高的增菌效率。研究结果表明,改良培养基配方和改善培养条件均可以有效缩短副溶血性弧菌检测的前增菌时间。大幅缩短增菌时间可以为实现食源性致病菌快检奠定基础。

In order to effectively shorten the pre-enrichment time and broken the technical bottleneck of rapid detection of Vibrio parahaemolyticus,this study explored a technical method to improve the preenrichment speed of Vibrio parahaemolyticus by optimizing the culture medium formula and culture conditions.The optimal medium formula obtained through single-factor and response surface analysis:soybean peptone 7.4 g/L,yeast extract 9.8 g/L,and bovine heart extract 20.6 g/L,NaCl 30.0 g/L,pH 8.0.The optimal culture conditions are:temperature 36 ℃,rotational speed 180 r/min.Applying the optimized pre-enrichment culture method,the Vibrio parahaemolyticus culture could be enriched within 6 hours to meet the sensitivity requirements for real-time polymerase chain reaction(PCR) detection.In samples with added food matrices,the optimized pre-enrichment culture method still showed high enrichment efficiency compared to the GB 4789.7.The results indicate that improving culture medium formula and culture conditions can effectively reduce the pre-enrichment time for V.parahaemolyticus detection,Significantly reducing the pre-enrichment time can lay a foundation for achieving rapid detection of foodborne pathogens.