食品科学 ›› 2010, Vol. 31 ›› Issue (20): 413-416.doi: 10.7506/spkx1002-6630-201020087

• 分析检测 • 上一篇    下一篇

Western 斑点印迹法技术检测致病性副溶血弧菌

杨靖亚,张 建,赵 勇,陶 妍,陆小凡,晁若瑜   

  1. 上海海洋大学食品学院
  • 收稿日期:2010-06-29 出版日期:2010-10-25 发布日期:2010-12-29
  • 通讯作者: 张建 E-mail:zhangjiancaiyun@sina.com
  • 基金资助:

    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)

Detection of Pathogenic Vibrio parahaemolyticus by Western Blotting

YANG Jing-ya,ZHANG Jian,ZHAO Yong,TAO Yan,LU Xiao-fan,CHAO Ruo-yu   

  1. College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China
  • Received:2010-06-29 Online:2010-10-25 Published:2010-12-29
  • Contact: ZHANG Jian E-mail:zhangjiancaiyun@sina.com

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。

关键词: 耐热直接溶血毒素, Western 斑点印迹, 致病性副溶血弧菌, 检测

Abstract:

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.

Key words: TDH;Western blotting, pathogenic Vibrio parahaemolyticu, detection

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