作　　者： ; ; ;

机构地区： 华南农业大学

出　　处： 《华南农业大学学报》 2011年第1期108-111,共4页

摘　　要： 将EMA（Ethidium monoazide）选择渗透性和PCR技术相结合,建立检验副溶血弧菌Vibrio parahaemolyticus死活细胞的EMA-PCR新方法.研究表明,25μg/mL或更高质量浓度的EMA可以抑制2.0×107CFU/mL的副溶血弧菌死细胞DNA的扩增,未经EMA处理的对应样品PCR反应呈阳性;而用相同浓度EMA处理2.0×107CFU/mL的活细胞菌悬液能扩增出目的产物.同时,对EMA处理中卤素灯的曝光时间进行优化,确定EMA处理的最佳曝光时间为5~10 min.结果证明,该方法是一种快速有效的检测死活细胞的新方法,能有效避免PCR检测假阳性结果的出现. A new method with the viable/dead stain ethidium monoazide（EMA） in combination with PCR（EMA-PCR） was developed.The results showed that the use of 25 μg/mL or more of EMA completely inhibit the PCR amplification of DNA derived from dead Vibrio parahaemolyticus cells containing 2.0×107 CFU/mL,but the samples without EMA treatment were PCR positive.The use of the same concentration of EMA did not inhibit the PCR amplification of DNA derived from viable bacteria.Meanwhile,the light exposure time to active and photolyze EMA was optimized,and the optimal light exposure time was 5-10 min.It is concluded that the EMA-PCR technique may provide an efficient new approach for distinction between the viable and dead cells and efficiently avoid false positive results.

关 键 词： 副溶血弧菌 死活细胞

领　　域： [轻工技术与工程] [轻工技术与工程]