摘要
目的:对副溶血弧菌(Vibrio parahaemolyticus,VP)的直接耐热溶血毒素(thermostable direct hemolysin,TDH)进行分离纯化,建立Western斑点印迹法技术检测致病性副溶血弧菌的方法。方法:用直接耐热溶血毒素免疫小鼠得到特异性抗血清,利用棋盘方法确定免疫血清最佳工作浓度,并对致病性与非致病性副溶血弧菌分别进行特异性测定。结果:最佳免疫血清最佳工作浓度为1:3200;检测致病性副溶血弧菌为阳性,非致病性副溶血弧菌属呈阴性。结论:Western斑点印迹法技术可以特异性检测致病性副溶血弧菌,灵敏度高、操作简单,可用肉眼判定结果。
Objective：To purify thermostable direct hemolysin （TDH） from the culture of Vibrio Parahaemolyticus and to establish western blotting method for determining pathogenic Vibrio parahaemolyticus. Methods：The specific anti-serum was obtained from immune mice using TDH. The working concentration of serum samples was determined by chessboard assay. The specificities of pathogenic Vibrio parahaemolyticus and non-pathogenic Vibrio parahaemolyticus were also determined. Results：The working concentration of serum samples was 1：3200. The samples with clear dots were judged as the positive reaction. V. parahaemolyticus was positive in pathogenic Vibrio parahaemolyticus,while non-pathogenic Vibrio parahaemolyticus was negative. Conclusion：Western blotting can be used to detect pathogenic Vibrio parahaemolyticus with the characteristics of high sensitivity,simple operation and eye-observable results.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2010年第20期413-416,共4页
Food Science
基金
上海市教育委员会科研创新项目(09YZ265)
上海市教育委员会重点学科建设项目(J50704)
