γ射线对副溶血性弧菌辐射效应的PCR检测

PCR Detection for the Irradiation Effects of γ-ray on Vibrio Parahaemolyticus

目的:研究γ射线对副溶血性弧菌(Vibrio parahaemolyticus)的辐射效应。方法:采用0kGy^7.0kGy内不同剂量的60Co-γ对浓度为2.0×108CFU/mL的副溶血性弧菌进行辐射,通过菌落计数计算该菌的致死率与存活率,并采用菌液PCR和常规PCR间接检测DNA浓度研究其致死效应。结果:菌落计数表明,在0kGy^3.0kGy内,该菌致死率随辐射剂量增加而呈线性增长;3.0kGy以上可全部杀灭试验浓度的副溶血性弧菌。常规PCR和菌液PCR结果表明,PCR扩增条带亮度都随着辐射剂量的增大而增强。常规PCR中,辐射剂量3.0kGy以上,电泳条带的亮度明显增加。结论:γ射线辐射剂量与存活副溶血性弧菌数、DNA的释放量都有明显的剂量-反应关系,辐射致死的主要原因是由射线对菌体细胞膜造成损伤、DNA外流造成的。

Objective：In order to study the irradiation effects of ^（60）Co γ-ray on Vibrio parahaemolyticus.Method：The Vibrio parahaemolyticus at the concentration of 2.0 ×108CFU/mL was irradiated by different doses of 60 Co γ-ray between 0 kGy～7.0 kGy.The lethality rate and survival were evaluated by colony-counting method,bacterium liquid PCR and conventional PCR were used to detect the DNA concentration indirectly.Result：The result of colony-counting method showed that The lethality rate raised linearly with increasing of 60 Co γ-ray dose.Vibrio parahaemolyticus can be killed completely by γ-ray irradiation at the dose of 3.0 kGy in the experiment concentration.The result of conventional PCR and bacterium liquid PCR showed that the brightness of PCR bands added with the increasing of ^（60）Co γ-ray doses.Above 3.0 kGy,the brightness of PCR bands were brighter than before.Conclusion：There was obvious dose-reaction relationship between the γ-ray irradiation dose and the CFU of survivals bacteria along with the leakage of DNA.The primary reason lead to the death of Vibrio parahaemolyticus might be the damage of cell membrane resulted from γ-ray of 60 Co and the leakage of DNA and protein.