大肠杆菌O157:H7胶体金试纸条研制  

Preparation of Colloidal Gold Strip for the Detection of Escherichia coli O157:H7

作　　者：黄岭芳[1] 段霞[1] 陈媛 刘文娟 魏华[1] 赖卫华[1] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,江西南昌330047 [2]江西中德生物工程有限公司,江西南昌330029

出　　处：《食品科学》2010年第24期355-359,共5页Food Science

基　　金：科技部中小企业创新基金项目(10C26223601934);南昌市重大产业化项目(2009)

摘　　要：采用胶体金标记抗大肠杆菌O157:H7单克隆抗体(鼠源),通过将大肠杆菌O157:H7多克隆抗体和驴抗鼠抗体(二抗)喷涂于硝酸纤维素膜分别作为检测线和质控线,研制大肠杆菌O157:H7胶体金快速检测试纸条。通过优化实验,确定最佳条件为标记量16.8μg/mL、标记pH8.0、封闭剂PEG20000、检测时样品最佳pH7.0～7.5。对26株常见细菌交叉反应结果表明,该试纸条除与鼠伤寒沙门氏菌ATCC13311和金黄色葡萄球菌CMCC26003有轻微交叉反应外,与其他24株菌均无交叉反应。该试纸条灵敏度为104CFU/mL。用该试纸条检测大肠杆菌O157:H7操作简便、快捷、灵敏度高、特异性强。Anti-Escherichia coli O157：H7 monoclonal antibody was labeled with colloidal gold to prepare a strip for the detection of E. coli O157： H7. Anti-E. coli O157：H7 polyclonal antibody and donkey anti-mouse IgG were dispensed on the nitrocellulose membrane as the test line and control line, respectively. The optimal conjugation was performed using 16.8μg/mL antibody at the condition of pH 8.0 and blocked with PEG20000. However, the optimal pH range for the detection was 7.0-7.5. Cross-reaction test was conducted by 26 bacteria strains. Results showed no obvious cross-reaction were observed except for the slight cross-reaction from Salmonella typhimurium ATCC13311 and Staphylococcus aureus CMCC26003. Meanwhile, the sensitivity of the developed strip was up to 104 CFU/mL. Moreover, this kind of developed strip was characteristics of simple operation, rapid detection, high sensitivity and specificity.

关 键 词：大肠杆菌O157:H7 胶体金 试纸条 

分 类 号：TS207.4[轻工技术与工程—食品科学]