呋喃西林代谢物荧光偏振免疫检测方法研究  

Determination of Nitrofurazone Metabolite Using Fluorescence Polarisation Immunoassay

作　　者：沈玉栋[1] 张世伟[1] 蔡肇婷[1] 雷红涛[1] 肖治理[1] 王弘[1] 孙远明[1] 

机构地区：[1]华南农业大学食品质量与安全研究所广东省食品质量与安全重点实验室,广东广州510642

出　　处：《分析测试学报》2009年第1期27-31,36,共6页Journal of Instrumental Analysis

基　　金：广东省自然科学基金资助项目(06300421;06300423);粤港关键领域重点突破项目(2006A25005002);高等学校博士学科点专项科研基金资助项目(20050564011)

摘　　要：利用新制备的抗体首次建立了呋喃西林代谢物氨基脲(SEM)的荧光偏振免疫分析(FPIA)方法。通过设计合成新的SEM半抗原CEPSEM(2-(4-((2-carbamoylhydrazono)methyl)phenoxy)aceticacid),偶联载体蛋白后免疫新西兰大白兔,制备了亲和力高、特异性好的多克隆抗体。结合新设计合成的荧光示踪物CEPSEM-HDF建立了SEM的FPIA方法。结果表明:在示踪物浓度为0.5nmol/L,抗体稀释度为1/100的优化条件下,IC50为47.9μg/L,检出限为8.3μg/L,线性范围为15.8～145.7μg/L。该方法特异性强(和其它相关兽药交叉反应小于0.1%),稳定性好(批内相对标准偏差小于2.5%,批间相对标准偏差小于6.3%)。A fluorescence polarisation immunoassay （FPIA）was developed for the determination of semicarbazide （SEM） via associating novel synthetic fluorescence tracer with novel prepared polyclonal antibody against 2- （2-nitrobenzylidene） hydrazinecarboxamide （ NPSEM ）. A novel and efficient hap- ten 2-（4-（（2-carbamoylhydrazono） methyl ） phenoxy ） acetic acid （ CEPSEM ） was designed, synthe- sized and coupled to the carrier protein BSA to give immune antigen. The animal immunization was used to produce a highly sensitive and specific polyclonal antibody. Under optimal conditions, the calibration curve showed a good linearity in the range of 15.8 - 145.7 μg/L. The detection limit of SEM was 8.3 μg/L and the ICs0 was 47.9 μg/L. The RSDs of intra and inter-assay of the method were less than 2.5% and 6.3% , respectively. The FPIA showed high stability, sensitivity, and specificity for the determination of SEM.

关 键 词：呋喃西林 示踪物 荧光偏振免疫分析(FPIA) 

分 类 号：R392-33[医药卫生—免疫学]