江西农业大学学报
江西農業大學學報
강서농업대학학보
ACTA AGRICULTURAE UNIVERSITATIS JIANGXIENSIS
2014年
3期
661-664
,共4页
丁丽军%张家禾%周作红%刘莉%洪伟鸣%孟婷%管远红%左伟勇
丁麗軍%張傢禾%週作紅%劉莉%洪偉鳴%孟婷%管遠紅%左偉勇
정려군%장가화%주작홍%류리%홍위명%맹정%관원홍%좌위용
恩诺沙星%残留%检测%活化酯法%间接竞争ELISA
恩諾沙星%殘留%檢測%活化酯法%間接競爭ELISA
은낙사성%잔류%검측%활화지법%간접경쟁ELISA
enrofloxacin%residue%detection%the active ester method%indirect competitive ELISA
通过活化酯法将小分子的恩诺沙星(EFLX)偶联到牛血清白蛋白(BSA)和鸡卵清白蛋白(OVA)上,形成偶联物EFLX-BSA和EFLX-OVA,以EFLX-BSA为免疫原试验小鼠,以EFLX-OVA为检测原通过ELISA检测小鼠血清效价,以及Ci-ELISA检测抗体的特异性和最低检测限。结果表明免疫原和检测原均成功合成,血清效价检测结果第一组为25600,第二组和第三组均大于51200,第二组最高;间接竞争ELISA法结果表明,所获抗体对EFLX特异性强,最低检出限为1.17 ng/mL,IC50值为54.60 ng/mL,检测范围为7.39~403.43 ng/mL。本试验所获得抗体效价高、特异性强,可以为ELISA试剂盒和胶体金试纸条研发提供良好的抗体材料。
通過活化酯法將小分子的恩諾沙星(EFLX)偶聯到牛血清白蛋白(BSA)和鷄卵清白蛋白(OVA)上,形成偶聯物EFLX-BSA和EFLX-OVA,以EFLX-BSA為免疫原試驗小鼠,以EFLX-OVA為檢測原通過ELISA檢測小鼠血清效價,以及Ci-ELISA檢測抗體的特異性和最低檢測限。結果錶明免疫原和檢測原均成功閤成,血清效價檢測結果第一組為25600,第二組和第三組均大于51200,第二組最高;間接競爭ELISA法結果錶明,所穫抗體對EFLX特異性彊,最低檢齣限為1.17 ng/mL,IC50值為54.60 ng/mL,檢測範圍為7.39~403.43 ng/mL。本試驗所穫得抗體效價高、特異性彊,可以為ELISA試劑盒和膠體金試紙條研髮提供良好的抗體材料。
통과활화지법장소분자적은낙사성(EFLX)우련도우혈청백단백(BSA)화계란청백단백(OVA)상,형성우련물EFLX-BSA화EFLX-OVA,이EFLX-BSA위면역원시험소서,이EFLX-OVA위검측원통과ELISA검측소서혈청효개,이급Ci-ELISA검측항체적특이성화최저검측한。결과표명면역원화검측원균성공합성,혈청효개검측결과제일조위25600,제이조화제삼조균대우51200,제이조최고;간접경쟁ELISA법결과표명,소획항체대EFLX특이성강,최저검출한위1.17 ng/mL,IC50치위54.60 ng/mL,검측범위위7.39~403.43 ng/mL。본시험소획득항체효개고、특이성강,가이위ELISA시제합화효체금시지조연발제공량호적항체재료。
Ovalbumin ( OVA) and bovine serum albumin ( BSA) were used as protein carriers to synthesize small molecule drugs EFLX complete antigens by the active ester method .The conjugates formed in this way were EFLX-BSA and EFLX-OVA.The serum titer was detected by means of EFLX-BSA immunogen and EFLX-OVA coating antigen by ELISA ,the specificity and detection limit ( LD) were detected by Ci-ELISA.The results showed that the immunogen and coating antigen were successfully synthesized .The serum titer detection result of the first group was 25 600,those of the second and third groups were more than 51 200,that of the second was the highest of all .The result of Ci-ELISA indicated that this method was sensitive and had a linear detec-tion range from 7.39 ng/mL to 403.43 ng/mL ( R2=0.993 9) ,with IC50 and LOD values of 54.60 ng/mL and 1.17 ng/mL.This experiment obtained high antibody titer and the specificity was strong ,which provides good antibody materials for the development of colloidal gold test and ELISA kit .