食品科学

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MTT法测定瑞士乳杆菌MB 2-1活菌数

杨培洁,李 腾,陈晓红,李 伟,董明盛   

  1. 南京农业大学食品科技学院,江苏 南京 210095
  • 出版日期:2013-10-25 发布日期:2013-09-28

MTT Colorimetric Method for Lactobacillus helveticus MB 2-1 Viable Cell Counting

YANG Pei-jie,LI Teng,CHEN Xiao-hong,LI Wei,DONG Ming-sheng   

  1. College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
  • Online:2013-10-25 Published:2013-09-28

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。

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

Abstract:

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.

Key words: MTT colorimetric method, Lactobacillus helveticus MB 2-1, viable cell counting, optimization