上转磷光免疫层析检测肠出血性大肠杆菌O157  

Development of UCP-immunochromatography Test for Rapid Detection of E.coli O157

作　　者：王静[1] 周蕾[1] 李伟[1] 胡孔新[1] 陈维娜[1] 闫中强[2] 杨瑞馥[2] 

机构地区：[1]中国检验检疫科学研究院卫生检疫研究所,北京100025 [2]军事医学科学院微生物流行病研究所,北京100071

出　　处：《中国食品卫生杂志》2007年第1期41-44,共4页Chinese Journal of Food Hygiene

基　　金：国家自然科学基金资助(30500455);国家质量监督检验检疫总局重大科研专项资助(2005IK0143)

摘　　要：目的建立一种肠出血性大肠杆菌O157的上转磷光免疫层析快速检测方法。方法利用上转换磷光标记和双抗体夹心免疫层析技术检测大肠杆菌O157,评价了该法的敏感性和特异性,并用于模拟污染食品样品的检测。结果该法能在40min内完成检测,应用于不同的肠杆菌科细菌如其他大肠杆菌、沙门菌、志贺菌、变形杆菌、产气肠杆菌、枸橼酸杆菌、沙雷菌、耶尔森菌以及葡萄球菌、副溶血弧菌、单增李斯特菌等23种28株常见细菌,未发现有交叉反应,检测灵敏度为5×103CFU/ml,每次最低可检测500个细菌,对奶粉、咖啡粉、饼干、蛋糕、绿豆糕、果冻、燕窝、果汁等样品中的人工染菌均可检测,最低检测浓度为5×103CFU/ml。结论肠出血性大肠杆菌O157上转磷光免疫层析方法简便快速,特异性和灵敏性好,适用于现场的快速检测。Objective To develop a method for rapid detection of Enterohemorrhagic E. coli O157 （EHEC O157） on the spot. Method An Up-Converting Phosphor （UCP） -immunoehromatography test （ICT） based on double-antibody sandwich assay was developed for detection of EHEC O157. The sensitivity and specificity of the method were evaluated. The feasibility of screening the bacteria in food samples were evaluated by different food samples artificially contaminated with EHEC O157 ： H7, including milk, starch, coffee, biscuit, cake, juice, etc. Results Typical analysis time was less than 40 minutes per sample. The sensitivity of the test was 5 ×10^3 CFU/ml. No cross reaction was found when the method was used to detect other Enterobacteriaceae （including E. coli, Salmonella, Shigella, Proteus, Morganella, Citrobacter, Enterobacter, Serratia ）, Staphylococus and V. parahemolyticus, confirming that the method is highly specific for E. coli O157. Conclusion The UCP- immunoehromatography test provides a rapid, specific and sensitive measure for detecting EHEC O157 on the spot.

关 键 词：大肠杆菌0157 发光 上转换发光材料 免疫学技术 

分 类 号：R155.5[医药卫生—营养与食品卫生学]