胶体金免疫层析法快速检测动物组织中残留的喹乙醇
Colloidal gold based on immunochromatographic assay for detection of olaquindox residues in animal tissues
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摘要: 研制胶体金免疫层析法试纸条,用于动物源食品中喹乙醇药物残留的快速检测。将已制备纯化过的喹乙醇多克隆抗体与胶体金结合产生的金标抗体喷涂在玻璃纤维垫上,并将包被抗原OLA-OVA和羊抗鼠二抗分别结合在硝酸纤维素膜上,组装成免疫层析快速检测试纸条,并检测试纸条的灵敏度、特异性、准确性和稳定性。将制备的试纸条用于检测猪肉和猪肝样品,结果表明本研究制备的试纸条可以在10min内完成检测,肉眼观察的最低检测限为0.05μg/m L,与其结构类似物卡巴氧、乙酰甲喹、3-甲基-喹噁啉-2-羟酸的反应交叉性小,该试纸条假阳性率小于5%,假阴性率为0,在干燥常温条件下保存6个月仍然有效。本实验研究的检测方法可应用于动物组织中喹乙醇的快速检测和现场筛查。Abstract: The aim of the present study was to develop colloidal gold immunochromatography assays for rapid detecting olaquindox residues in animal origin food. Combining the prepared and purified polyclonal antibody of olaquindox with colloidal gold and sprayed on glass fibre mat. Coatingen ( OLA- OVA) and sheep anti- mouse Ig were fixed on nitrocellulose membrane, and the immunochromatography testing paper strip was then manufactured, to detect its sensitivity, specificity, accuracy and stability. The test strip was used for detection of olaquindox residues in pork and pig livers.The results showed that the test strip could be accomplished qualitative detection of olaquindox in 10 minutes, and the limit of detection of the strips was 0.05μg / m L in both pork and pig livers.There was very little cross- reactivity detected with carbadox, mequindox and 3- methyl- quinoxaline- 2-hydroxy acids.The false positive rate was less than 5%, and the false negative rate was 0, and no effectiveness losing occurred during 6months of storage at room temperature under dry conditions.These results suggested that the prepared strip can be applied in rapid detection and on- site screening of olaquindox residues in animal tissues.