Western 斑点印迹法技术检测致病性副溶血弧菌

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。

Detection of Pathogenic Vibrio parahaemolyticus by Western Blotting

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.