链霉素残留检测ELISA试剂盒的研制及应用  

Development and Application of Competitive ELISA Kit for Rapid Detection of Streptomycin Residues

作　　者：范国英[1] 王顺岗[1] 王自良[1] 张改平[2] 邓润广[2] 杨艳艳[2] 

机构地区：[1]河南科技学院动物科学学院,新乡453003 [2]河南省农业科学院免疫学重点实验室,郑州450002

出　　处：《畜牧兽医学报》2008年第3期380-384,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：“十一五”国家科技支撑计划重大项目(2006BAK02A21)

摘　　要：本研究采用EDC法人工合成链霉素抗原（SM—BSA），免疫BALB／c小鼠，通过杂交瘤技术获得了分泌抗链霉素单克隆抗体的杂交瘤细胞株，建立了快速检测链霉素（SM）残留的酶联免疫方法，并成功研制了试剂盒。然后对其灵敏度、准确度、特异性、基质效应性等技术指标进行检测，同时应用该试剂盒对生长猪尿液中的残留情况进行检测。结果表明：除双氢链霉素外，该试剂盒与其它抗生素类药物均无交叉反应；线性检测范围为1～1289g／L，相关系数R^2=0．9251，灵敏度为0．45μg／L，半数抑制浓度（IC50）为7．769g／L，检测限为19g／L；猪尿样的平均添加回收率为84．1％；基质对该试剂盒的检测结果影响不大。该试剂盒具有快速、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 of streptomycin（SM-Kit）was developed with anti-SM monoclonal antibody（SM mAb） and its sensitivity, veracity, specificity and the effect of matrix were tested, respectively. At the same time, the urine of the swine was detected with this ELISA-Kit. The results indicated that the SM-Kit has little or no cross-reactivity with other anti-biotics besides of dihydrostreptomycin;the linear detection was 1.0 to 128.0 μg/L（R^2=0. 925 1）, the sensitivity was 0.45 μg/L,the IC50 was 7.76 μg/L and the detection limit was 1 μg/L. The recoveries of SM spiked in pig urine was 84.1%. The matrix has no effect on the results of the SM-Kit. The SM-Kit possessed rapidity, sensitivity, specificity and briefness and was proved useful in the rapid test of SM residues in animal.

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]