氨苯磺胺免疫荧光分析方法的建立  

Establishment of Immunofluorescent Assay for Sulfanilamide

作　　者：李龙[1] 沈红[1] 黄园园[1] 陈静[1] 

机构地区：[1]北京农学院动物科学技术系,北京102206

出　　处：《动物医学进展》2007年第8期17-20,共4页Progress In Veterinary Medicine

基　　金：北京市教委资助(200410020029);北京市优秀人才培养专项经费资助(20041D0502106)

摘　　要：通过重氮化法将SN与人血清白蛋白(HSA)偶联制备全抗原SN-HSA,用全抗原免疫新西兰大白兔,制备了抗SN的多克隆抗体(AbSN),AbSN与温度敏感水凝胶偶联形成水凝胶复合物(pNIPA-AbSN),全抗原(AgSN)与异硫氰基荧光素偶联形成荧光复合物(FITC-AgSN)。FITC-AgSN和游离半抗原SN竞争性地与有限量的水凝胶复合物pNIPA-AbSN反应。根据水凝胶相变温度以上免疫复合物沉淀的特性进行分离,结果表明,该法对SN测定的线性范围在1 ng/mL～100μg/mL,50%抑制率时检出限为60.45 ng/mL。　A novel immunoassay for sulfanilamide（SN） was established.SN was diazotizated and coupled to human serum albumin（HSA） to form a complete antigen（AgSN） for animal immune and preparing its polyclonal antibody（AbSN）.Then the AbSN was conjugated to poly-N-isopropylacrylamide（pNIPA） to produce a hydrogel antibody conjugate（pNIPA-AbSN）.The complete antigen was labeled by a fluorescein isothiocyanate（FITC） to form the fluorescent antigen complex（FITC-AgSN）.A competitive immunofluorescent assay method based on the competition of FITC-AgSN and free SN with limited amounts of pNIPA-AbSN was developed.The separation in the immunological reaction was achieved by the precipitation of the immunocomplex above pNIPA critical temperature.The results showed that the calibration range was 1 ng/mL100 μg/mL and the determination level for SN in term of 50% inhibition rate was 60.45 ng/mL.

关 键 词：免疫分析 氨苯磺胺 水凝胶 

分 类 号：S859.84[农业科学—临床兽医学]