原料乳中β-内酰胺酶检测试剂盒的研究

Study on the test kit of β-lactamase in raw milk

中文关键词:  β-内酰胺酶  试剂盒  乳制品

英文关键词:β-lactamase  test kit  dairy products

中文摘要:

      目的 制备原料乳中β-内酰胺酶的快速检测试剂盒, 方法 β-内酰胺酶分解青霉素钾的产物青霉噻唑酸具有还原性, 能将二价铜还原为一价铜, 2, 2′-联喹啉与一价铜反应生成的是紫红色配合物, 颜色深浅与β-内酰胺酶的量在一定范围内呈线性关系。 结果 利用可见分光光度法优化出试剂盒最佳检测条件为: 0.05% 2, 2′-联喹啉添加量为2.4 mL, 20 mmol/L的CuSO4最佳添加量0.2 mL, 原料乳添加量为0.1 mL; 配制β-内酰胺酶的标准品浓度梯度溶液0～200 U/mL进行显色反应后, 利用分光测色仪测定颜色反应L*、a*和b*值数据, 并应用色彩模块软件对其进行模拟, 制作标准比色卡; 原料乳样品显色15 min后与标准比色卡对比,确定牛奶中β-内酰胺酶浓度, 试剂盒最低检出限为4 U/mL。结论 β-内酰胺酶检测试剂盒法具有快速、操作简单、经济实用等特点, 易于普及和现场测定。

英文摘要:

      Objective To make the kit for rapid detection of β-lactamase in raw milk. Methods Penicilloic acid, the decomposer of penicillin G potassium by β-lactamase, has the reducibility and can reduce Cu2+ into Cu ， the 2, 2'-biquinolyl can coupling with Cu to display purple, which the color depth and the amount of β-lactamase are in linearity within a certain range. Results Using visible spectrophotometry to optimize the best detection condition of box for: 0.05% 2, 2'-Alliance quinoline added volume for 2.4 mL, 20 mmol/L of CuSO4 for 0.2 mL, raw dairy samples added volume for 0.1 mL; β-Lactamase standard solution for gradient concentration 0~200 U/mL for colour reaction is prepared, using spectro colorimeter to determine the color reaction L*, and a* and b* value data, and applicate the color module software on its for simulation to prepare the standard color card. After 15 min for Color rendering of sample, it is compared to the standard color cards to judge the concentrations of β-lactamase in a sample. The detection limit of Kit was 4 U/mL. Conclusion β-lactamase kit method is of rapid, simple, economical and practical, easy to popularize and field tests.