Establishment of Single Tube Quantitative Detection Technology for Escherichia coli Assay Based on Fluorescent Method

Abstract

通过研究培养温度、培养基pH值对大肠杆菌生长和培养液荧光强度的影响,确定44℃,培养液pH值7.0~7.5为大肠杆菌MUG酶荧光检测的最佳条件。通过研究培养时间和接种菌浓度与培养液荧光强度的关系,建立起基于荧光方法检测大肠杆菌的单管定量检测技术。通过与平板菌落计数法和最大可能数法比较发现单管定量检测法的相对标准偏差小于这两种常用的方法,而且更为快速、经济,适合用于大肠杆菌的定量检测。Influence of incubation temperatures and pH values of culture liquid on growth of Escherichia coli and the fluorescent intensity of culture liquid had been investigated.E.coli incubated at 44℃ in the culture liquid with a pH value in the range of 7.0~7.5 was found to be the best condition for fluorescent assay.Single tube technology to detect E.coli was established through the studies on the relationship between incubation time,inoculation quantity of E.coli and the fluorescent intensity of culture liquid.Relatively standard diversity of single tube detection technology was lower than those of colony-counting method and the most probably number method.The results indicated that the single tube quantitative detection method was fit for E.coli quantitative detection because of the several advantages such as less time-consuming and more economical compared with traditional methods.国家高技术研究发展计划项目(863计划项目)(No2002AA639600);; 福建省自然科学基金(NoB0410022

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