荧光光度法快速定量大肠杆菌的实验方法研究

Study on the rapid enumeration of Escherichia coli with fluorescent method

研究比较了菌体渗透处理方式与培养方式对大肠杆菌荧光光度法检测效果的影响,以及考察了培养条件对荧光强度的影响,结果表明仅采用渗透处理方式处理菌液会造成荧光检测值偏小,灵敏度偏低;但经LST-MUG(pH7.0)培养液44℃培养7h后的荧光值能对100.9～106.8cells/mL范围内的大肠杆菌进行定量,结果与国标方法无显著性差异。该法进一步缩短了大肠杆菌的检测时间至8h左右,且重现性良好,适合于大肠杆菌的定量检测。

The effect of osmotic treatment and culture on the detection of E.coli with fluorescent method were discussed respectively,and the effect of culture conditions on fluorescent intensity also investigated.The results indicated that only osmotic treatment for E.coli would cause small fluorescent intensity and low detection sensitivity.However,fluorescent intensity obtained under the conditions of cultured for 7h at 44℃ in the LST-MUG culture medium could quantified the E.coli in the range of 100.9～106.8cells/mL,and the results obtained by this method and standard method showed no significant difference.Optimized fluorescent method shortened the detection time to 8h,and showed good reproducibility.Therefore,the method described is applicable to the rapid enumeration of E.coli.