免疫磁分离结合胶体金免疫层析法快速检测大肠杆菌O157∶H7  

Establishment of a Method Combined Immunomagnetic Separation with Colloidal Gold Lateral Flow Assay and Its Application in Rapid Detection of Escherichia. coli O157∶H7

作　　者：崔希[1] 熊齐荣[1,2] 熊勇华[1] 山珊[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,南昌330047 [2]中国检验检疫科学研究院食品安全研究所,北京100123

出　　处：《分析化学》2013年第12期1812-1816,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.31271863);南昌大学食品科学与技术国家重点实验室自由探索课题(No.SKLF-ZZB-201307);南昌市科技项目(No.2012-CYH-DW-SP-001)资助

摘　　要：以抗大肠杆菌O157∶H7单克隆抗体偶联磁珠制得免疫磁珠;以胶体金标记的抗大肠杆菌O157∶H7单克隆抗体为标记抗体,抗大肠杆菌O157∶H7多克隆抗体和驴抗鼠二抗分别作为检测线和质控线成分制备胶体金免疫层析试纸条。将免疫磁珠分离和胶体金免疫层析法相结合应用于大肠杆菌O157∶H7的现场快速检测中。结果表明:1 mL 7.6×103CFU/mL菌液经过免疫磁珠分离富集后,用胶体金免疫层析法检测可得到阳性结果。对于10 CFU/g大肠杆菌O157∶H7污染的食品样本,经过6 h预增菌,免疫磁珠分离和胶体金试纸条检测72 min即可检出。本方法的建立对于食品中有害物质的现场快速检测具有重要意义。The immunomagnetic beads were prepared by coupling anti-Escherichia, coli O157：H7 monoclonal antibody with magnetic beads. The colloidal gold test strip was composited with anti-O157：H7 monoclonal antibody marked by colloidal gold as the detection antibody, anti-E, coli 0157： H7 polyclonal antibody as the test line and the donkey anti-mouse IgG secondary antibody as the control line. Immunomagnetic separation was combined with colloidal gold lateral flow assay, and this method was applied in the rapid detection of E. coli O157 ：H7. The results showed that 1 mL of 7.6×10^3 CFU/mL of E. coli O157：H7 gave a positive result by using colloidal gold lateral flow after immunomagnetic separation. Food sample spiked with 10 CFU/g of E. coli O157 ： H7 after 6 h of enrichment could be detected within 72 min of immunomagnetic separation and colloidal gold lateral flow assay. The method is of great value to the rapid on-site detection of harmful substance in food.

关 键 词：免疫磁分离 胶体金 免疫层析 大肠杆菌O157 H7 

分 类 号：R155[医药卫生—营养与食品卫生学] O657.1[医药卫生—公共卫生与预防医学]