MTT法测定大肠杆菌活菌数实验研究.docVIP

第31卷第12期2011年12月 环境科学学报Acta Scientiae Circumstantiae Vol．31,No．12Dec．,2011 基金项目:天津市应用基础及前沿技术研究计划项目(No．11JCZDJC25000 Supported by the Tianjin Research Program of Application Foundation and Advanced Technology (No．11JCZDJC25000作者简介:汪志荣(1965—,女,教授(博士, E-mail :wangzr@tjut．edu．cn ;*通讯作者(责任作者Biography :WANG Zhirong (1965—,female ,professor (Ph．D．,E-mail :wangzr@tjut．edu．cn ;*Corresponding author 汪志荣,高琼,马传鑫,等．2011．MTT 法测定大肠杆菌活菌数实验研究[J ]．环境科学学报, 31(12:2642-2650Wang Z R ,Gao Q ,Ma C X ,et al ．2011．Experimental study on the application of MTT colorimetric method in counting live Escherichia coli [J ]．Acta Scientiae Circumstantiae ,31(12:2642-2650 MTT 法测定大肠杆菌活菌数实验研究 汪志荣* ,高琼,马传鑫,李燕妮天津理工大学环境科学与安全工程学院,天津300384收稿日期:2011-02-25 修回日期:2011-03-27 录用日期:2011-04-12 摘要:实验分析MTT 比色法中的工作波长、反应时间、MTT 用量、菌龄、二甲基亚砜(DMSO 等因素对大肠杆菌活细菌计数的影响．同时,通过与平板计数对比,确定了MTT 细菌计数的可行性．结果表明,当细菌数在107 109cell ·mL －1范围内时,双波长MTT 法可用于检测大肠杆菌的活菌数量,最佳反应时间为60min ,MTT 溶液用量200μL ,溶解剂DMSO 用量1mL ,且该方法对菌龄不敏感．OD 510－690值与细菌浓度C 呈良好的正向线性关系,其可决系数在0．99以上．关键词:MTT ;双波长测定;大肠杆菌;活菌计数文章编号:0253-2468(201112-2642-09中图分类号:X172 文献标识码:A Experimental study on the application of MTT colorimetric method in counting live Escherichia coli WANG Zhirong *,GAO Qiong ,MA Chuanxin ,LI Yanni School of Environmental Science and Safety Engineering ,Tianjin University of Technology ,Tianjin 300384Received 25February 2011; received in revised form 27March 2011; accepted 12April 2011 Abstract :Experimental studies were conducted on the influences of working wavelength ,reaction range of microbial cell number ,reaction time ,concentration of MTT ,cell age of Escherichia coli (E．coli ,DMSO in the MTT colorimetric method (MTT method ,etc．for determining live E．coli．Compared with dilution －plate method ,the MTT method was a viable method for counting live E．coli ,provided the number of bacteria was in the range 107cell ·mL －1 109cell ·mL －1．The best working parameters were reaction time 60minutes ,MTT 200μL and DMSO 1mL．The method was insensitive to cell age of E．col