| 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1～128 μg/L,相关系数R2=0.925 1,灵敏度为0.45μg/L,半数抑制浓度(IC50)为7.76 μg/L,检测限为1 μg/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。 |
| 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 |
| DOI： |
| 分类号: |
| 基金项目:国家“863”计划项目（2001AA249030） |
|
| Filtration of hybridoma lines and development of rapid test competitive ELISA kit for streptomycin |
|
|
| Abstract: |
| 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（R2＝0.925 1），the sensitivity of 0.45 μg/L,the IC50 of 7.76 μg/L and the detection limit of 0.45 μg/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. |
| Key words: Streptomycin,monoclonal antibody,cell fusion,competitive ELISA,rapid test kit |
