竞争化学发光酶免疫检测动物源性食品中四环素残留
Determination of Tetracycline Residues in Animal Derived Foods by Competitive Chemiluminescent Enzyme Immunoassay
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摘要: 目的:建立一种准确度高、重复性好的检测四环素(tetracycline,TC)残留的竞争化学发光酶免疫测定法(competitive chemiluminescence enzyme immunity,CLEIA)。方法:通过选择适宜的偶联载体蛋白,优化反应温度、时间等条件,建立了一种测定四环素的竞争化学发光酶免疫测定法(TC-CLEIA)。结果:该方法的检测范围为0.0259~95.9478 μg/L,相关系数r为0.9998,检测限为0.0133 μg/L,准确度为99.17%,灵敏度为0.7305 μg/L,变异系数<10%,对氯霉素、青霉素具有极低的交叉反应;向空白猪肉、鱼肉、鸡肝中添加三种不同浓度的四环素标准品时,检测结果具有较高的准确度和重复性;建立的TC-CLEIA测定法检测了四环素小鼠中毒模型中肝脏、肾脏和肌肉中四环素残留的浓度,浓度大小依次为:肾脏 > 肝脏 > 肌肉。结论:本研究建立的TC-CLEIA检测法简便,检测限低,准确度、重复性、特异性均很好,适用于动物源性食品中四环素残留的检测。Abstract: Objective:A great accurate and repeatable method for the determination of tetracycline(TC)by competitive chemiluminescence enzyme immunoassay assay(CLEIA)was established. Method:The TC-CLEIA,which was used to detect the concentration of TC,was established by choosing appropriate coupling carrier,optimizing reaction temperature and reaction time. Results:The detection range of TC-CLEIA for detection TC was 0.0259~95.9478 μg/L with 0.9998 of r. And the detective limit was 0.0133 μg/L,accuracy was 99.17%,sensitivity was 0.7305 μg/L and variation coefficient was less than 10%. There was little cross-reaction with chloramphenicol and penicillin in TC-CLEIA. The detection results had higher accuracy and repeatability when the different concentration of TC standard was added into the blank pork,fish and liver of chicken. The concentrations of TC residues in liver,kidney and muscle detected by TC-CLEIA using tetracycline-toxic mice model. The results showed that the order of concentration was kidney > liver > muscle. Conclusion:The TC-CLEIA method with high sensitivity and specificity was suitable for the detection of TC residues in animal-derived foods.
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