NADH荧光法快速检测细菌总数

Rapid Detection of Total Bacterial Number Using ADH Fluorescence Method

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1 nmol/L, NADH含量在10 nmol/L~0.2 mmol/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 342 nm为激发波长, 461 nm为发射波长测定提取液荧光强度, 1 h内可检测到样品1×104 CFU/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。

Abstract:

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(R2= 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 1×104 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.