食品科学 ›› 2007, Vol. 28 ›› Issue (2): 238-240.

• 分析检测 • 上一篇    下一篇

两步荧光法检测牛乳中大肠杆菌的研究

 付丽娜, 刘宁, 霍贵成   

  1. 东北农业大学乳品科学教育部重点实验室; 东北农业大学乳品科学教育部重点实验室 黑龙江哈尔滨150030; 黑龙江哈尔滨150030;
  • 出版日期:2007-02-15 发布日期:2011-12-31

Study on E.coli Detection in Cowmilk by Two-Step Fluorescent Method

 FU  Li-Na, LIU  Ning, HUO  Gui-Cheng   

  1. Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China
  • Online:2007-02-15 Published:2011-12-31

摘要: 本文主要进行了两步荧光检测牛乳中大肠杆菌的研究,确定了两步荧光法的基本条件和过程。其过程为将大肠杆菌菌悬液在TB培养基中进行12h的预培养后,取出0.1ml菌液加入到1.5ml的离心管中,再加入0.2mg/mlMUG底物0.1ml和100μg/ml的多粘菌素B0.05ml培养45min,最后加入pH10.4的0.2mol/LGly-NaOH缓冲液1.75ml终止反应,使用荧光分光光度计测荧光密度。空白样为0.1mlmol/LUG底物加0.05ml100ug/ml的多粘菌素B,再加0.2mol/LpH10.4Gly-NaOH缓冲液1.75ml。

关键词: 两步荧光法, 检测, 大肠杆菌

Abstract: The assay conditions and procedure were studied in this paper. Firstly, Escherichia coli was pre-incubated in TB for 12 hours. Then 0.1ml aliquots were taken to be combined with 0.05ml polymyxin B(100μg/ml) and 0.1ml MUG(4-methylum belli feryl-β-D-glucuro nide)(0.2mg/ml) and incubated at 37℃ for 45min. Finaly 1.75ml Gly-NaOH buffer(0.2mol/L, pH10.4) were added to end the reaction. The the two-step fluorescent method is better than that of MPN(most probable number), which is widely used for the detection of Escherichia coli,and was quantitatively close to the traditional MPN method.

Key words: two-step fluorescent method, detection, Escherichia coli