饲料中喹乙醇ELISA检测试剂盒的研制及性能分析

doi:10.11843/j.issn.0366-6964.2013.08.018

Abstract

Abstract:

The aim of this study was to develop a rapid detection kit for olaquindox (OLA-Kit) in animal feed. The OLA monoclonal antibody (OLA mAb) was applied to assemble the indirect competitive ELISA kit and its characterization was determined. The results showed that the calibration curve of OLA-Kit was a typical sigmoid curve with the linear detection of 1-243 μg· L^－1, and R² was 0.976 7. The kit has a half inhibition concentration (IC₅₀) of (14.29±3.75) μg·L^－1, the detection limit of 0.79 μg·L^-1, the quantitative limit of 4.0 μg·L^－1, the cut-off value of 83.37 μg·g－1 for pannage, and the matrix has no effect on the results of the OLA-Kit; The recoveries of OLA in pig feed, chicken feed and fish feed were all above 74.40%, and the inter-assay and intra-assay coefficient variation were all below 15%; The OLA-Kit had 17.93% cross-reactivity towards Kuianchun and little or no cross-reactivity towards MQCA, QCA and other quinoxaline drugs. The validity of kit in 2-8 ℃ was above six months. There was no significant difference of the recovery of OLA in pig feed between OLA-Kit and HPLC (P≥0.05). In conclusion, the OLA-Kit was sensitive, specific and accurate, could be used for the rapid determination and extensive screening of OLA in animal feed.

CLC Number:

S859.796

ZHANG Jing-yan, WANG Lei, LI Jian-xi, ZHANG Hong, ZHANG Kai, YANG Zhi-qiang, WANG Xue-zhi, QIN Zhe, MENG Jia-ren. Development and Performance of ELISA Kit for Rapid Detection of Olaquindox in Feed[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(8): 1297-1304.

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References

［1］朱学加，陈金兰，何计国.喹乙醇人工抗原的合成及鉴定［J］.食品科学，2008，29（5）：129-133.

［2］董尧，王前勇.喹乙醇的应用研究进展［J］.畜牧兽医杂志，2008，27（3）：63-65.

［3］朱柱振，陈杖榴，冯淇辉.喹乙醇在鸡体内的药物动力学及组织浓度研究［J］.畜牧兽医学报，1993，24（3)：259-264.

［4］高洪，陈莲，杨建发，等.喹乙醇致鸡中毒后线粒体膜的ATPase活性变化研究［J］.西南农业学报, 2007，20（5)：1120-1123.

［5］郝利华, 肖希龙, 周宗灿.穿梭质粒pSP189/ 哺乳动物Vero细胞检测系统在喹乙醇诱变分子机制研究中的应用［J］.癌变·畸变·突变, 2006, 18（1）：23-25.

［6］中华人民共和国农业部第168号公告［S］.饲料药物添加剂使用规范，2001.

［7］王永红，丁平，朱英才.饲料中喹乙醇检测方法研究［J］.南方农业，2011，5（9)：51-52.

［8］GIZZI G, VINCENT U, VON HOLST C, et al. Validation of an analytical method for the determination of carbadox and olaquindox in feedstuff by liquid chromatography coupled to UV and/or diode array detection［J］. Food Addit Contam，2007，24（11）：1226-1235.

［9］HU K, YANG X L. Determining olaquindox in fish tissue & feed by improved HPLC method(high performance liquid chromatography)［J］.J Shanghai Fish Univ，2003，12：28-32.

［10］曾静，朱宽正，王鹏.高效液相色谱-串联质谱法测定水产品中的喹乙醇［J］.中国食品卫生杂志，2006，18（5）: 423- 425.

［11］朱立平，陈学清.免疫学常用实验方法［M］.北京：人民军医出版社，2000.

［12］杨利国，胡少旭，魏平华，等.酶联免疫测定技术［M］.南京：南京大学出版社，1985.

［13］王磊.喹乙醇ELISA检测技术单克隆抗体制备与评价［D］.北京:中国农业科学院，2012.

［14］农医发［2005］17号农业部文件.关于发布“兽药残留酶联免疫试剂（盒）备案审查技术资料要求”和“兽药残留酶联免疫试剂（盒）备案参考评判标准”的通知［S］.2005.

［15］中华人民共和国国家标准.饲料中喹乙醇的测定-高效液相色谱法［S］.GB/T 8381.7-2009.

［16］中国兽药典委员会.兽药使用指南［M］.化学药品卷.北京：中国农业出版社，2006.

［17］张改平，刘庆堂，邓瑞广，等.磺胺二甲基嘧啶残留快速检测阻断ELISA试剂盒的研制及其性能测定［J］.中国预防兽医学报，2009，31（1)：51-55.

［18］唐伟国.医学检验诊断试剂的制备与应用［M］.上海：科学技术文献出版社，1996.

［19］范国英，王顺岗，王自良，等.链霉素残留检测ELISA试剂盒的研制及应用［J］.畜牧兽医学报，2008，39（3）: 380-384.

［20］宋春美，李青梅，刘庆堂，等.喹乙醇单克隆抗体的制备及其免疫学特性的鉴定［J］.核农学报，2010，24（4）: 777-783.

［21］刘智宏，叶妮，黄耀凌，等.ELISA法测定牛奶和鸡肌肉中四环素类药物残留［J］.中国兽药杂志，2006，10（11）: 1-4.

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Development and Performance of ELISA Kit for Rapid Detection of Olaquindox in Feed

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