氯霉素残留检测阻断ELISA试剂盒的研制及性能测定  

Development and evaluation of a rapid test blocking ELISA kit for chloramphenicol residues

作　　者：杨艳艳[1] 张改平[1] 邓瑞广[1] 王自良[2] 王选年[2] 柴书军[1] 邢广旭[1] 

机构地区：[1]河南省动物免疫学重点实验室,河南郑州450002 [2]河南科技学院动物科学学院,河南新乡453003

出　　处：《中国预防兽医学报》2007年第2期130-134,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家863项目"快速检测试纸条技术在动物疫病和食品残留检测中的研究及应用"(2001AA249030)资助

摘　　要：在建立氯霉素单克隆抗体(CAP mAb)杂交瘤细胞株和阻断ELISA方法的基础上,研制出CAP残留快速检测试剂盒(CAP-kit),并对其性能进行了测定。结果表明,CAP-kit的标准曲线呈典型的S型,符合4参数logit曲线拟合,相关系数R2=0.9953,检测范围为1.0μg/L～128.0μg/L,灵敏度为0.85μg/L,半数抑制浓度(IC50)为7.54μg/L,检测限为1.0μg/L;牛奶样、猪肉样的平均添加回收率为88.3%、82.5%,平均批内和批间变异系数均<15%;CAP-kit与氯霉素琥珀酸钠的交叉反应(CR%)为150%,与其它酰胺醇类和抗菌素药物无交叉反应;试剂盒在4℃可保存6个月。An ELISA kit for detection of chloramphenicol（CAP-kit）was developed using a CAP monoclonal antibodies（CAP mAb）, which was produced from hybridoma lines. The kit was evaluated for its accuracy, specificity and reliability. The calibration curve of CAP-kit was shown to be a typical sigmoid curve fitted to the four parameters logistic equation with the linear detection of 1.0 to 128.0 μg/L（R^2=0.9953）. It has a sensitivity of 0.85 μg/L, the IC500 of 7.54 μg/L and the detection limit of 1.0 μg/L. The recoveries of CAP spiked in milk and in pork were 88.3 % and 82.5 % respectively. The inter-assay and intra-assay coefficient variation was shown below 15 %. The CAP-kit had 150 % cross-reactivity with CAP succinate sodium and little or no cross-reactivity with other amphenicols. At 4 ℃, this kit remained effective for more than six months.

关 键 词：氯霉素 单克隆抗体 阻断ELISA 快速检测试剂盒 

分 类 号：S854.4[农业科学—临床兽医学]