改良Vero细胞毒性试验法检测志贺毒素  

The measurement of Shiga toxin by modified Vero cell cytotoxicity assay

作　　者：毕旺来[1] 杨俊[1] 杜文芳[1] 李睿[1] 

机构地区：[1]武汉轻工大学生物与制药工程学院,湖北武汉430023

出　　处：《武汉轻工大学学报》2017年第2期22-25,共4页Journal of Wuhan Polytechnic University

摘　　要：采用Vero细胞培养法检测5株肠出血性大肠杆菌O157食品分离株的志贺毒素。在48孔细胞培养板中培养Vero细胞24 h,然后加入梯度稀释的志贺毒素滤液。培养24 h及48h后在倒置显微镜下观察Vero细胞形态。培养48 h后结晶紫染色,测定Vero细胞半致死时,对应的志贺毒素滤液稀释倍数。证实形态观察和结晶紫染色相结合可以准确测定肠出血性大肠杆菌O157食品分离株的志贺毒素。In this study,titers of Shiga toxins of five Enterohemorrhagic Escherichia coli O157 strains isolated from food have been measured by modified Vero cell cytotoxicity assay. Vero cells were incubated plate,and then mixed with serial diluted Shiga toxin filtrates. After 24h and 48h incubation,the cell morphologywas observed under an inverted microscope. After 48h incubation, the Vero cells wand the toxin titers were determined as the reciprocal of the highest dilution of filtrates that caused 50% Vero celldeath. This study demonstrated that morjDhology observation in combination witli crure the Shiga toxin produced by Enterliemorrhagic Escherichia coli O157 strains isolated from food.

关 键 词：细胞培养 肠出血性大肠杆菌O157 志贺毒素 Vero细胞毒性 

分 类 号：R155.5[医药卫生—营养与食品卫生学]