草鱼肠型点状产气单胞菌双抗夹心ELISA试剂盒的研制

通过对包被单抗和酶标单抗以及抗原最佳工作浓度的优化,建立草鱼肠型点状产气单胞菌双抗夹心ELISA试剂盒检测体系。用分光光度法对检测体系的特异性、稳定性和灵敏度进行测定;用脱脂奶平板法对试剂盒的临床应用进行检测。结果表明,单抗最适稀释液为pH9.4的碳酸盐缓冲液,洗涤液为含φ=0.05%Tween-20的PBS,包被单抗最适体积稀释倍数为1∶800,酶标单抗最适体积稀释倍数为1∶6 000,抗原最佳工作浓度为3×104cfu/mL。试剂盒与肠型点状产气单胞菌有特异性反应,脱脂奶平板法与试剂盒法对草鱼患病检出率基本一致,符合率达90%;试剂盒有很好的储存稳定性,检测灵敏度为7×103cfu/mL。

Development of Double antibody Sandwich ELISA Kit for Aeromonas punctata fintestinalis Ctenopharyngodon Idellus

Double-antibody sandwich ELISA kit for Aeromonas punctata fintestinalis Ctenopharyngodon idellus was developed on the base of establishing double-antibody sandwich ELISA method,determining working concentration of coating monoclonal antibody and enzyme-labeling monoclonal antibody,as well as the best working concentration of antigen;measuring specificity,stability and sensitivity of the kit by spectrophotometry,and detecting the clinical application of the kit by skim milk plates.We used PBS(pH=7.4) as dilu...