用试纸法快速检测β-半乳糖苷酶活性的研究

Rapid Detection of Beta-galactosidase Activity by Test Paper

应用5-溴-4-氯-3-吲哚-β-半乳糖苷(X-gal)试纸建立一种快速检测β-半乳糖苷酶活性的方法。其原理在于酶色原底物X-gal在β-半乳糖苷酶作用下产生蓝色的5-溴-4-氯-3-吲哚。X-gal的价格昂贵,常规法每个平皿的X-gal的用量为3～4mg。该试验以大肠杆菌为阳性对照菌株,无乳链球菌为阴性对照菌株,用试纸法对新疆石河子周边地区牛场分离出的101株链球菌是否产β-半乳糖苷酶进行检测。共筛选出阳性菌21株,检出率为20.79%;阴性菌80株,检出率为79.21%。采用试纸法不仅使X-gal用量大大减少,节省成本,同时操作快速、简便、准确。

With 5-bromo-4-chloro-3-indole-beta-galactosidase （X-gal） test paper,a rapid method for detection of beta-galactosidase activity was developed.Its principle is that the chromogen substrate X-gal is catalyzed by beta-galactosidase to produce blue 5-bromo4-chloro-3-indole.X-gal is very expensive,and each plate needs 3～4 mg X-gal for conventional methods.The developed test paper method was used to detect 101 Streptococcus strains isolated form cattle breeding farms around Shihezi City,with E.coli as positive control and Streptococcus agalactiae as negative control.A total of 21 strains produced beta-galactosidase,and the positive rate was 20.79%.The other strains did not produce beta-galactosidase,and the negative rate was 79.21%.The developed test paper method needs much less X-gal and reduces cost.The operation is also rapid,simple and accurate.