食品科学 ›› 2011, Vol. 32 ›› Issue (2 ): 333-337.doi: 10.7506/spkx1002-6630-201102077

• 技术应用 • 上一篇    

食品中四环素类残留的酶联免疫检测试剂盒的研制

国占宝1,武玉香2,田文礼1,赵亚周1,彭文君1,*,柳家鹏2   

  1. 1.中国农业科学院蜜蜂研究所,农业部授粉昆虫生物学重点开放实验室,国家蜂产品加工技术研发分中心,北京      100093;2.北京华安麦科生物技术有限公司,北京      102200
  • 收稿日期:2010-09-16 出版日期:2011-01-25 发布日期:2011-01-10
  • 通讯作者: 彭文君 E-mail:pengwenjun@vip.sina.com
  • 基金资助:

    科技部科研院所技术开发专项(2009EG134274);国家现代蜜蜂产业技术体系项目(NYCYTI-43-KXJ17);农业部“948”项目(2006-G19)

Development of A New ELISA Kit for Tetracycline Residue Detection in Foods

GUO Zhan-bao,WU Yu-xiang,TIAN Wen-li,ZHAO Ya-zhou,PENG Wen-jun,LIU Jia-peng   

  1. 1. Institute of Apiculture Research, Chinese Academy of Agricultural Sciences, Key Laboratory for Pollinating Insect Biology, Ministry of Agriculture, National Research and Development Center For Bee Product Processing, Beijing      100093, China;
    2. Huaan Magnech Bio-tech Co. Ltd., Beijing      102200, China
  • Received:2010-09-16 Online:2011-01-25 Published:2011-01-10

摘要: 目的:基于定量检测动物源性食品中四环素类抗生素的残留量,开发研制四环素类抗生素竞争酶联免疫(ELISA)检测试剂盒。方法:采用杂交瘤技术,筛选并克隆稳定分泌抗四环素(CEL)的杂交瘤细胞株,利用免疫BALB/c 小白鼠制备抗四环素单克隆抗体。结果:logit/log 拟合标准曲线为y= - 2.3x + 2.79,相关系数r=0.9971,线性检测范围为0.05~4.05ng/mL,半数抑制剂量(IC50)为0.293ng/mL,灵敏度为0.05ng/mL,检测限为3~5ng/mL;组织、牛奶、蜂蜜的添加回收率分别为(90 ± 10)%、(85 ± 10)%、(90 ± 10)%;平均批内和批间变异系数均小于15%;四环素与金霉素等同类抗生素的交叉反应率大于60%,与其他类抗生素无交叉反应;此试剂盒在4℃可保存180d 以上。结论:建立的试剂盒是符合技术指标的要求,可用于动物源性食品中四环素类抗生素残留的定量检测。

关键词: 食品, 四环素, 单克隆抗体, 竞争酶联免疫

Abstract: A competitive ELISA kit for tetracycline (TC) residues detection was developed by employing TC monoclonal antibody prepared by hybridoma technology using anti-TC monoclonal antibodies secreted by immunized BALB/c mouse. Performance tests showed that the logit/log calibration curve of the kit was y = -2.3x + 2.79 (correlation coefficient, r = 0.9971), with a linear range from 0.05 to 4.05 ng/mL, the half inhibitory concentration (IC50) was 0.293 ng/mL, the sensitivity was 0.05 ng/mL and the detection limit ranged from 3 to 5 ng/mL. The recovery for spiked TC in muscle, milk, and honey was (90 ± 10)%, ( 85 ± 10)% and (90 ± 10)%, with coefficient of variations (CV%) less than 15%, respectively. Cross-reactivity between TC and CTC was more than 60%, and there was no cross-reactivity between TC and other antibiotics. This ELISA Kit could be stored at 4 ℃ for more than six months.

Key words: food, tetracycline, monoclonal antibodies, competitive ELISA

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