荧光法快速检测细菌总数.pdfVIP

微生物学通报 MAY 20, 2009, 36(5): 773~779 Microbiology © 2009 by Institute of Microbiology, CAS tongbao@ 生物实验室 NADH 荧光法快速检测细菌总数 * 王 晶 王静雪 林 洪 梅册霞 ( 266003) 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 4 CFU/mL 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 ×10 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。 关键词: NADH, 荧光法, 细菌总数, 快速检测 Rapid Detection of Total Bacterial Number Using NADH Fluorescence Method * WANG Jing WANG Jing-Xue LIN Hong MEI Ce-Xia (Food Science and Engineering Department, Ocean University of China, Qingdao, Shandong 266003, China) 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 re- sult 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