克伦特罗和莱克多巴胺多残留胶体金免疫层析试纸条的研制

畜牧兽医学报

克伦特罗和莱克多巴胺多残留胶体金免疫层析试纸条的研制

姜金庆^1,2，杨雪峰¹，王自良^1*，邓瑞广²，王选年²，张改平^2*

(1. 河南科技学院动物科学学院，新乡 453003； 2. 河南省动物免疫学重点实验室，郑州 453003)

收稿日期:2012-05-28 出版日期:2013-01-23 发布日期:2013-01-23
通讯作者: 张改平， E-mail: zhanggaiping2003@126.com；王自良，E-mail: wangzl_2008@yahoo.com.cn
作者简介:姜金庆 (1973-)，男，河南方城人，副教授，博士，主要从事动物源食品安全免疫学快速检测方法研究，Tel：0373-3040079，E-mail: jjq5678@126.com
基金资助:
“十一五”国家科技支撑计划重大项目（2006BAK02A21/1）；河南省教育厅科技攻关项目资助（2011A230003）

Development of a Monoclonal Antibody Based Colloidal Gold Immunochromatography Strip for Multianalysis of Clenbuterol and Ractopamine

JIANG Jin-qing^1,2, YANG Xue-feng¹, WANG Zi-liang^1*, DENG Rui-guang², WANG Xuan-nian², ZHANG Gai-ping^2*

(1. College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China; 2. Henan Provincial Key Laboratory of Animal Immunology, Zhengzhou 453003, China)

Received:2012-05-28 Online:2013-01-23 Published:2013-01-23

摘要/Abstract

摘要：

本试验旨在研制同时检测克伦特罗（CL）和莱克多巴胺（RAC）的多残留胶体金免疫层析试纸条（Multi-strip）。用细胞融合技术筛选CL和RAC杂交瘤细胞株,制备单克隆抗体,金标抗体竞争性膜层析技术研制Multi-strip。该试纸条由样品垫、偶联垫、双检测线和质控线标记的NC膜、吸收垫等几部分构成，其中偶联垫上灌注有CL和RAC两种金标单克隆抗体，双检测线由检测抗原CL-OVA和RAC-BSA喷膜构成，间距2 mm。结果表明：多残留试纸条的CL和RAC目测检测限分别为1和2 ng·mL^－1，检测时间5~8 min。基质呈阳性的真实样品用单残留试纸条、ELISA和GC-MS检测，结果一致，无假阳性或假阴性案例。该试纸条具有快速、敏感、特异、简便等特点，适合于现场检测和筛选。

Abstract:

The objective of this study was to develop a colloidal gold-labeled multianalysis strip assay (Multi-strip) for the simultaneous detection of clenbuterol (CL) and ractopamine (RAC) residues. Hybridoma lines that secrete monoclonal antibody (mAb) against CL and RAC were filtered through cell fusion technology and their immunological traits were characterized. The colloidal gold-based immunochromatography competition method was employed to develop a Multi-strip, which consisted of a sample pad, a conjugate pad, a NC membrane containing two test lines and a control line, and an absorbent pad. The gold-labelled CL mAb and RAC mAb were sprayed onto the conjugate pad, and the test lines were separately coated with CL-OVA and RAC-BSA, 2 mm in distance. The visual detection limits of Multi-strip were 1 and 2 ng·mL^－1 for CL and RAC, respectively, and the results can be judged within 5-8 min. Parallel analysis of natural positive matrices with CL and RAC showed consistent results obtained from the individual strip, ELISA and GC-MS, without false-positive or false-negative cases. This Multi-strip possesses rapidity, sensitivity, specificity and simplicity, and is suitable for on-site screening and qualitative determination.

中图分类号:

S859.84

姜金庆，杨雪峰，王自良，邓瑞广，王选年，张改平. 克伦特罗和莱克多巴胺多残留胶体金免疫层析试纸条的研制[J]. 畜牧兽医学报, doi: .

JIANG Jin-qing, YANG Xue-feng, WANG Zi-liang, DENG Rui-guang, WANG Xuan-nian, ZHANG Gai-ping. Development of a Monoclonal Antibody Based Colloidal Gold Immunochromatography Strip for Multianalysis of Clenbuterol and Ractopamine[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/

https://www.xmsyxb.com/CN/Y2013/V44/I1/87

参考文献

［1］申屠芬琴，强致懿，张素霞，等. 猪肾脏等组织中盐酸莱克多巴胺的HPLC检测方法及其残留消除［J］. 畜牧兽医学报, 2007, 38(6):589-594.

［2］张海棠，王自良，邓瑞广，等. 抗莱克多巴胺单克隆抗体杂交瘤细胞株的建立及其免疫学特性鉴定［J］. 食品科学, 2009, 30(3):175-179.

［3］TURBERG M P, RODEWALD J M, COLEMAN M R. Determination of ractopamine in monkey plasma and swine serum by high-performance liquid chromatography with electrochemical detection ［J］. J Chromatogr B Biomed Appl, 1996, 675(2):279-285.

［4］PESSER A C, DICKSON L C, MACNEIL J K, et al. Determination of betaagonists in liver and retina by liquid chromatography-tandem mass spectrometry ［J］. J NOAC Int, 2005, 88(1):61-69.

［5］应永飞，皮雄娥，吴平谷，等. 气相色谱-质谱法同时测定动物尿样中莱克多巴胺和克伦特罗［J］. 质谱学报, 2006, 27(2):74-78.

［6］XU T, WANG B M, SHENG W, et al. Application of an enzyme-linked immunosorbent assay for the detection of clenbuterol residues in swine urine and feeds ［J］. J Environ Sci Health B, 2007, 42(2):173-177.

［7］ZHANG G P, WANG X N, YANG J F, et al. Development of an immunochromatographic lateral flow test strip for detection of β-adrenergic agonist clenbuterol residues ［J］. J Immunol Methods, 2006, 312(1-2):27-33.

［8］LI X, ZHANG G P, DENG Y G, et al. Development of rapid immunoassays for the detection of ractopamine in swine urine ［J］. Food Addit Contam, 2010, 27(8):1096-1103.

［9］WANG Z, ZHU Y, DING S, et al. Development of a monoclonal antibody-based broadspecificity ELISA for fluoroquinolone antibiotics in foods and molecular modeling studies of cross-reactive compounds ［J］. Anal Chem, 2007, 79(12):4471-4483.

［10］ZHU Y, LI L, WANG Z, et al. Development of an immunochromatography strip for the rapid detection of 12 fluoroquinolones in chicken muscle and liver ［J］. J Agric Food Chem, 2008, 56(14):5469-5474.

［11］SHIM W B, DZANTIEV B B, EREMIN S A, et al. One-step simultaneous immunochromatographic strip test for multianalysis of ochratoxin A and zearalenone ［J］. J Microbiol Biotechnol, 2009, 19(1):83-92.

［12］JIANG J Q, ZHANG H T, FAN G Y, et al. Preparation of monoclonal antibody based indirect competitive ELISA for detecting 19-nortestosterone residue ［J］. Chinese Sci Bull, 2011, 56(25):2698-2705.

［13］JIANG J Q, WANG Z L, ZHANG H T, et al. Monoclonal antibody-based ELISA and colloidal gold immunoassay for detecting NT residue in animal tissue ［J］. J Agric Food Chem, 2011, 59(18):9763-9769.

[1]	黄婧洁, 李苗, 陈莹娴, 钟雅兰, 张婷婷, 姜廷超男, 李建成. 基于免疫磁珠净化间接竞争酶联免疫吸附试验检测氟喹诺酮类药物[J]. 畜牧兽医学报, 2023, 54(2): 766-778.
[2]	王子琳, 唐子恒, 林泽宁, 刘文字, 刘戎, 张嘉慧, 沈祥广. 中药成分辛弗林对胶体金免疫层析法检测β-受体激动剂的影响[J]. 畜牧兽医学报, 2021, 52(12): 3675-3680.
[3]	陈莹娴, 马月姣, 李建成. 抗球虫药物残留的免疫检测技术[J]. 畜牧兽医学报, 2021, 52(4): 899-908.
[4]	曹玉颖, 王杰, 张希墨, 吴琴尧, 邱基程, 李淑媛, 张璐, 龚晓会, 白如念, 曹兴元. 盐酸多西环素片在羔羊体内残留消除规律及休药期分析[J]. 畜牧兽医学报, 2021, 52(4): 1086-1093.
[5]	王双, 杨海峰, 余波, 李士洋, 郑育基, 卜仕金. 烯丙孕素口服液在猪体内的残留消除研究[J]. 畜牧兽医学报, 2018, 49(9): 2005-2014.
[6]	张静，周倩，唐梦君，张小燕，赵敏，杨星星，万玉，高玉时. 疫苗油佐剂在鸡蛋和组织中的残留代谢分析[J]. 畜牧兽医学报, 2017, 48(3): 544-551.
[7]	高金芳,岳婷婷，赵祎,陈冬梅,袁宗辉. 兽药多残留样品前处理技术研究进展[J]. 畜牧兽医学报, 2017, 48(1): 8-22.
[8]	孙礼瑞，谢星，谢恺舟，张小杰，王剑锋，崔璐璐，刘建宇，张跟喜，戴国俊，王金玉. 氨苄西林在鸡组织中的残留消除规律[J]. 畜牧兽医学报, 2015, 46(1): 130-137.
[9]	李帅鹏，黄显会，孔祥凯，怀彬彬，王伟，王辉. 肌注马波沙星在猪组织中残留消除规律[J]. 畜牧兽医学报, 2014, 45(5): 827-832.
[10]	程古月，彭娟，王玉莲，董小冰，彭大鹏，郝海红，黄玲利，袁宗辉. β-内酰胺类抗生素残留受体分析法的研究进展[J]. 畜牧兽医学报, 2014, 45(3): 354-362.
[11]	李帅鹏，黄显会，孔祥凯，张晓会，李向阳. 头孢噻呋在猪组织中残留消除规律[J]. 畜牧兽医学报, 2013, 44(8): 1311-1316.