菌落PCR方法的建立及其与常规PCR方法的比较

Establishment of Colony PCR Method and Its Comparison with Conventional PCR Method

为建立一种快速、经济、敏感的PCR检测方法 ,以直接检测出环境、海产品等中的靶基因 ,通过PCR扩增已知携带tlh和tdh基因的副溶血弧菌标准菌株 ,并和常规PCR方法比较 ,初步建立了菌落PCR技术 ;再以副溶血弧菌标准菌株为阳性对照 ,以本实验室自海洋环境中分离得到的数株野生菌株为检测对象 ,采用新建立的菌落PCR以及常规PCR方法对它们进行扩增比较 ,进一步证明了菌落PCR技术检测携带相关基因菌株的可靠性 .通过比较 ,发现菌落PCR和常规PCR的结果相当一致 。

To establish a quick, sensitive and economical PCR method for directly detecting tlh and adh genes-carrying Vibrio parahaemolyticus in environment and seafood, a colony PCR method was developed and its results were compared with those of the conventional PCR method, thus establishing a colony PCR method. By using the Vibrio parahaemolyticus strains as positive controls, several marine bacteria strains preserved in the lab were amplified through the colony PCR method and the conventional PCR method, by which the reliability of colony PCR method for the corresponding gene detection was proved. The comparison shows that the results obtained by colony PCR method are in great agreement with those obtained by the conventional PCR method, and that the tlh and tdh genes in positive controls can be amplified by these two PCR methods.