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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 establish a rapid detection method for up-conversion phosphorylation immunochromatography of enterohemorrhagic Escherichia coli O157. Methods The Escherichia coli O157 was detected by up-conversion phosphorescence labeling and double-antibody sandwich immunochromatography. The sensitivity and specificity of this method were evaluated and used to test the contamination of contaminated food samples. Results The method can be completed within 40min detection, applied to different Enterobacteriaceae such as other Escherichia coli, Salmonella, Shigella, Proteus, Enterobacter aerogenes, Citrobacter, Serratia, Yersin Bacteria and Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes and other 23 kinds of 28 kinds of common bacteria found no cross-reaction, detection sensitivity of 5 × 103CFU / ml, each time a minimum of 500 bacteria can be detected on the milk powder, Artificial bacteria in samples such as coffee powder, biscuits, cakes, mung bean cakes, jelly, bird’s nest and fruit juice can be detected with the lowest detection concentration of 5 × 10 3 CFU / ml. Conclusion The transglutaminase immunoassay of enterohemorrhagic Escherichia coli O157 is simple, rapid, specific and sensitive and suitable for rapid detection in the field.