抗链霉素杂交瘤细胞株的建立与竞争ELISA试剂盒的研制  

Filtration of hybridoma lines and development of rapid test competitive ELISA kit for streptomycin

作　　者：范国英[1] 王建华[1] 朱金凤[2] 张改平[3] 邓瑞广[3] 刘庆堂[3] 杨继飞[3] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]河南农业职业学院,河南郑州451450 [3]河南省农业科学院免疫学重点实验室,河南郑州450002

出　　处：《西北农林科技大学学报（自然科学版）》2007年第4期46-50,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"863"计划项目(2001AA249030)

摘　　要：制备牛血清白蛋白-链霉素(SM-BSA)免疫Balb／c小鼠,用细胞融合技术制备抗链霉索单克隆抗体(SM mAb)杂交瘤细胞,体内诱生腹水法制备SM mAb。依据竞争ELISA原理,应用SM mAb研制SM快速ELISA检测试剂盒(SM-Kit),并对其灵敏度、准确度、特异性、基质效应性等进行检测。结果表明,SM-Kit标准曲线呈典型的S型,线性检测范围为1～128μg／L,相关系数R2=0．925 1,灵敏度为0．45μg／L,半数抑制浓度(IC50)为7．76μg／L,检测限为1μg／L;奶样、猪尿样的平均添加回收率分别为104．45％和84．1％;SM-Kit与双氢链霉素的交叉反应率(CR)为104．16％,与其他抗生素类药物的CR<0．001％;基质对SM-Kit的检测结果影响不大。可见SM-Kit具有快速、敏感、特异、简便等特点,适合于SM残留的快速检测,具有较高的推广应用价值。Balb/C mice were immunized with streptomycin-bovine serum albumin and hybridoma lines that secrete monoclonal antibody against streptomycin（SM mAb） were filtered with cell fusion. A competitive ELISA test kit for detection streptomycin（SM-Kit）was developed with anti-SM monoclonal antibody （SM mAb） and its sensitivity, veracity, specificity and stability were tested respectively. The calibration curve of the SM-Kit with standard SM competitor was typical sigmoid curve with the linear detection of 1.0 to 128.0 μg/L（R^2 =0. 925 1） ,the sensitivity of 0.45μg/L,the IC50 of 7.76 μg/L and the detection limit of 0.45μg/L. The recoveries of SM spiked in milk,pig urine were 104.45%,84.1% respectively. The SM-Kit had 104.16% cross-reactivity towards dihydrostreptomycin and little or no cross-reactivity towards other antibiotics. The SM-Kit possessed rapidity, sensitivity, specificity and briefness and was proved to be used in the rapid test of SM residues in animal food.

关 键 词：链霉素 单克隆抗体 细胞融合 竞争ELISA 快速检测试剂盒 

分 类 号：S859.84[农业科学—临床兽医学]