克伦特罗残留检测免疫芯片的研制

doi:10.3969/j.issn.1000-1158.2015.06.23

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摘要为实现克伦特罗残留的规模检测设计其残留检测的免疫芯片。以玻片为载体并对玻片表面进行APTES-DGA化学修饰，将克伦特罗抗体偶联至玻片表面, 以牛血清白蛋白封闭玻片表面活性基团，加待测品和标样，标样中辣根过氧化酶标记的克伦特罗和待测品中的克伦特罗与克伦特罗抗体竞争性结合，借助辣根过氧化酶催化化学发光反应并检测光信号值，检测过程中考查加样体积、温育温度、温育时间、洗液浓度及化学发光积分时间等对检测结果的影响，检测方法学试验中考查检出限、定量限、线性和回收率等指标。结果显示：通过APTES-DGA修饰载玻片具有较好的信号均一性和抗体偶联能力，检测过程中加样体积为50 μL、温育温度为37 ℃、温育时间为30 min、洗液浓度为10倍稀释液、化学发光积分时间为60 s具有较稳定的检测结果。本法研制的免疫芯片检测克伦特罗残留快速、简单、准确。

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	孙来玉
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	严琼琼
	陆云华
	杨金田

关键词 ：计量学, 免疫芯片, 药物残留检测, 克伦特罗, 化学发光

Abstract：Immunochip is designed in order to realize the massive detection of clenbuterol residue.Slide glass surface as the carrier is modified by APTES-DGA.The clenbuterol antibody is coupled to the slide glass surface.Glass surface active groups are blocked by bovine serum albumin. The samples and standards, horseradish peroxidase labeled clenbuterol are added into the immunochip.CLB and CLB-HRP are combined competitively with Ab (CLB).Chemiluminescence reaction is catalyzed by the horseradish peroxidase and optical signal value is detected Sampling volume, incubation temperature, incubation time, concentration of washing solution, chemiluminescence integral time are tested.The results show that glass slide with APTESDGA has a better signal uniformity and antibody coupling ability. In the detection process, immunochip signal is stable when sampling volume is 50 μL, incubation temperature is 37 ℃, incubation time is 30 min, wash solution concentration is 10 fold diluent and chemiluminescence reader integral time is 60 s. The method is fast, simple and accurate by immunochip analysis for clenbuterol residue.

Key words： metrology immunochip drug residue detection； clenbuterol chemiluminescence

PACS:

TB99

基金资助:浙江省公益性应用研究计划（分析测试）项目(2012C37027)

作者简介: 孙来玉（1973-），男，安徽无为人，湖州师范学院实验师，主要研究方向为生物医药与食品安全。sunly@hutc.zj.cn

引用本文:

孙来玉,钱坤,邵朝纲,尹莉,严琼琼,陆云华,杨金田. 克伦特罗残留检测免疫芯片的研制[J]. 计量学报, 2015, 36(6): 657-661.
SUN Lai-yu,QIAN Kun,SHAO Chao-gang,YIN Li,YAN Qiong-qiong, LU Yun-hua,YANG Jin-tian. Development of Immunochip for Detection of Clenbuterol Residue. Acta Metrologica Sinica, 2015, 36(6): 657-661.

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http://jlxb.china-csm.org:81/Jwk_jlxb/CN/10.3969/j.issn.1000-1158.2015.06.23 或 http://jlxb.china-csm.org:81/Jwk_jlxb/CN/Y2015/V36/I6/657

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