NADH荧光法快速检测细菌总数  

Rapid Detection of Total Bacterial Number Using NADH Fluorescence Method

作　　者：王晶[1] 王静雪[1] 林洪[1] 梅册霞[1] 

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003

出　　处：《微生物学通报》2009年第5期773-779,共7页Microbiology China

基　　金：国家自然科学基金(No.30500384);国家863计划项目(No.2006AA09Z431)

摘　　要：基于细菌胞内NADH的荧光特性及其在胞内含量稳定的特性,建立一种快速检测细菌总数的新方法。该荧光法的NADH检测限为1nmol/L,NADH含量在10nmol/L～0.2mmol/L间与荧光强度呈良好线性关系(R2=0.9905)。经离心获得菌体细胞,热Tris-HCl法提取胞内NADH,以342nm为激发波长,461nm为发射波长测定提取液荧光强度,1h内可检测到样品1×104CFU/mL菌数。结果表明该方法快速、灵敏、简便、重复性好,可适用于食品卫生与安全、环境检测等领域活细菌数量的定量检测。To set up a new method of detecting bacterial number in situ, NADH fluorescence method based on the fluorescent characteristic of NADH was used. When the concentration of NADH ranged from 10 nmol/L to 0.2 mmol/L, its concentration had a good line relationship to the fluorescence intensity（R^2= 0.9905）. Separating bacterial cells by centrifugation and extracting NADH with hot Tris-HCl buffer, the result of bacterial count detected with NADH standard plot was 1×10^4 CFU/mL in an hour. In summary, NADH fluorescence method is rapid, sensitive, simple and reliable to detect total bacterial number. Therefore, the method can be widely applied in the field of food sanitation and safety, environment detection and so on.

关 键 词：NADH 荧光法 细菌总数 快速检测 

分 类 号：Q93-3[生物学—微生物学]