摘要
目的建立CAP间接和直接ELISA检测方法。方法用重氮化法分别偶联氯霉素(Chloramphenicol,CAP)与BSA和OVA作为免疫原和检测原,腹腔注射8周健康雌BALB/c小鼠大量制备抗CAP的单克隆抗体并优化条件,建立了CAP的快速、灵敏、简便的直接和间接竞争ELISA方法。结果两种检测方法的回归方程和相关系数分别为:y=-25.411x+97.041,R2=0.9889和y=-27.768x+103.91,R2=0.9889,线性范围分别为1.87-75 ng/mL和0.93-25ng/mL,对CAP的最低检出浓度为0.97 ng/mL和0.35 ng/mL。并利用所建立的两种ELISA方法对鸡肉模拟样品进行了检测,其检测样品的回收率分别为98.607%,85.152%。结论为下步CAP的试纸条或试剂盒建立提供基础。
Objective To describe and contrast the direct and indirect competitive enzyme immunoassay for the quantitative analysis of CAP.Methods The ascites of CAP was prepared by intraperitoneal injection the hybridoma immunized by the diazotizated artificial antigen CAP-OVA to BALB/c mice,the indirect competitive inhibition ELISA and the direct competitive inhibition ELISA were applied to contrast.Results The regression equation and coefficient correlation of two methods were y=-25.411x+97.041,R2=0.9889 and y=-27.768x+103.91,R2=0.9889 respectively;the concentration limit to CAP were 0.97 ng/mL and 0.35 ng/mL.The assay carried out to detect the imitation sample using the two ELISA,the recovery rate were 98.607% and 85.152%.Conclusion This research would be useful for the development of anti-CAP EIA Kit or test paper.
出处
《中国实验诊断学》
2007年第11期1530-1533,共4页
Chinese Journal of Laboratory Diagnosis
基金
国家自然基金资助项目(30671762)
