糖皮质激素ELISA 检测方法的建立及群选性抗体的筛选

doi:10.7506/spkx1002-6630-200924073

食品科学 ›› 2009, Vol. 30 ›› Issue (24 ): 331-336.doi: 10.7506/spkx1002-6630-200924073

糖皮质激素ELISA 检测方法的建立及群选性抗体的筛选

胡拥明，王利兵，袁媛，陈伟，边爱，马伟，胥传来*

江南大学食品学院

收稿日期:2009-07-08 出版日期:2009-12-15 发布日期:2010-12-29
通讯作者: 胥传来* E-mail:xcl@jiangnan.edu.cn

Development of an ELISA for Detection of Dexamethasone and Screening of Polyclonal Antibody

HU Yong-ming，WANG Li-bing，YUAN Yuan，CHEN Wei，BIAN Ai，MA Wei，XU Chuan-lai*

School of Food Science and Technology, Jiangnan University, Wuxi 214122, China

Received:2009-07-08 Online:2009-12-15 Published:2010-12-29
Contact: XU Chuan-lai* E-mail:xcl@jiangnan.edu.cn

摘要/Abstract

摘要：

设计间接竞争ELISA 方法，并优化二抗稀释比、竞争时间、PBS 溶液的pH 值3 个条件参数。优化后的地塞米松、倍他米松、双氟米松3 种抗体的间接竞争ELISA 曲线的IC50 分别为2.66、7.36、6.12ng/ml。通过3 种抗体的交叉反应来评估其群选特性。结果表明，地塞米松抗体的群选性最强，对双氟米松的交叉反应率高达120%、倍他米松73%、曲安息龙37%、泼尼松龙21%，而对其他3 种内源性类糖皮质激素交叉反应率均小于0.1%，然后地塞米松以0.375、0.75、1.5、3.0ng/g 分别添加到鸡肉样本中测定回收率，所得回收率在61.3%～80.3% 之间，变异系数小于15%。在实际样本检测中，将ELISA 方法的测定结果与LC-MS 的结果进行比较，两者具有很好的相关性。

关键词: 糖皮质激素, 酶联免疫吸附测定法, 检测, 群选性抗体, 选择性

Abstract:

Indirect competitive ELISA methods were developed based on the high-titer antibody and the dilution ratio of goat anti-rabbit-HRP, competition time and the pH of PBS solution were optimized. The IC50 of optimized indirect compete ELISA for dexamethasone, betamethasone and flumethasone were 2.66, 7.36, 6.12 ng/ml, respectively. The group selection of three antibodies was accessed by detecting their cross-reactivities. The results showed that the anti-dexamethasone antiboidy was the best suitable for group selection and the cross-reactivity toward flumethasone, betamethasone, riacinolone and predinisolon was up to 120%, 73%, 37%, predinisolone 21% while the cross activity towards several natural glucocorticoids was lower than 0.1%. The recoveries were detected by spiking four concentrations (0.375, 0.75, 1.5 ng/ml and 3.0 ng/ml) of DEX in chicken muscle tissue and the results indicated that the recoveries were between 61.3% and 80.3% and the coefficients of variation were lower than 15%. The ELISA method and LC-MS method were applied to analyze the real samples, and high correlation was observed between the two methods.

Key words: corticosteroids, ELISA, detection, class specific antibody, group-selection

中图分类号:

R977.11

胡拥明，王利兵，袁媛，陈伟，边爱，马伟，胥传来*. 糖皮质激素ELISA 检测方法的建立及群选性抗体的筛选[J]. 食品科学, 2009, 30(24 ): 331-336.

HU Yong-ming，WANG Li-bing，YUAN Yuan，CHEN Wei，BIAN Ai，MA Wei，XU Chuan-lai*. Development of an ELISA for Detection of Dexamethasone and Screening of Polyclonal Antibody[J]. FOOD SCIENCE, 2009, 30(24 ): 331-336.

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糖皮质激素ELISA 检测方法的建立及群选性抗体的筛选

Development of an ELISA for Detection of Dexamethasone and Screening of Polyclonal Antibody

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