MTT法测定瑞士乳杆菌MB2-1活菌数  

MTT Colorimetric Method for Lactobacillus helveticus MB2-1 Viable Cell Counting

作　　者：杨培洁[1] 李腾[1] 陈晓红[1] 李伟[1] 董明盛[1] 

机构地区：[1]南京农业大学食品科技学院,江苏南京210095

出　　处：《食品科学》2013年第20期99-102,共4页Food Science

基　　金：国家自然科学基金面上项目(31371807);国家自然科学基金青年科学基金项目(31201422);国家"863"计划项目(2011AA100903;2013BAD18B01-4);国家农业科技成果转化资金项目(2012GB23600639);江苏省自然科学基金项目(BK2011651);教育部博士点基金新教师类课题(20110097120028);江苏高校优势学科建设工程资助项目(PAPD)

摘　　要：以赛里木拉丝酸奶中提取的瑞士乳杆菌MB 2-1(Lactobacillus helveticus MB 2-1)为研究对象,探讨MTT法用于细菌计数的可行性及测量细菌数量的范围,优化MTT法用于瑞士乳杆菌活菌计数的工作波长、MTT和DMSO用量、反应时间。结果表明:在107～109CFU/mL内测出的OD值与细菌浓度呈正向线性关系(R2>0.99),最佳反应条件为:1.0mL待测菌液与0.2mL MTT溶液在40℃条件下染色反应1h,反应液需要经过12000r/min离心10min,取离心沉淀物溶解于1.0mL二甲基亚砜中,于工作波长510nm处测定此溶液的OD值。MTT比色分析法可用于检测乳酸菌活菌数量。The feasibility of MTT colorimetric method for Lactobacillus helveticus MB2-1 viable cell counting was evaluated, and the wavelength, MTT dosage, DMSO dosage and incubation time were optimized. Results showed that the OD value was linear with the cell concentration （R2 〉 0.99） at cell concentrations between 107 CFU/mL and 109 CFU/mL. The best working procedure was described as following： 1.0 mL of bacterial suspension reacted with 0.2 mL of MTT solution for 1 h min at 40 ℃. After centrifugation at 12000 r/min for 10 min, 1.0 mL of DMSO was added to dissolve the precipitate. OD510nm values were measured. This study indicates that the MTT colorimetric method can be applied for lactobacillus viable cell counting.

关 键 词：MTT染色计数法 瑞士乳杆菌MB 2-1 活菌计数 条件优化 

分 类 号：TS201.3[轻工技术与工程—食品科学]