摘要
目的:建立基于重组溶血素单抗的单核李斯菌ELISA检测方法。方法:从5株抗单核增生性李斯特菌的杂交瘤细胞中,筛选亲和力较高的单克隆抗体,用W estern-b lot鉴定、Prote in A纯化、HRP标记,经交叉配伍筛选出最佳的抗体组合。结果:建立了单核李斯特菌的ELISA检测方法,对单核增生性李斯特菌的最低检出浓度为5.2×108/L,本检测方法特异性强,与常见细菌没有交叉反应。结论:基于重组蛋白单抗的单核李斯菌ELISA检测方法的建立为单核李斯特菌的特异性诊断提供了有效手段。
Objective：To establish a recombinant listeriolysin O Monoclonal antibody Based ELISA for detection of Listeria monocytogenes.Methods： Five strains of Monoclonal Antibodies were obtained by traditional methods and purified with protein A column,identificated By Western Blot,labed with HRP.Couple of Monoclonal antibodies for coating and labing separately was selected from 5 Strains of hybridoma cells.Results： A sandwich ELISA for detection of rLLO and Listeria monocytogenes was established with a concentration limit of 1.95 ng/ml rLLO and 5.2×108/L Listeria monocytogenes respectively.This method had a high specificity and had no cross reaction with other bacteria.Conclusion： rLLO monoclonal antibody based sandwich ELISA for Listeria monocytogenes.Detection would have practical importance for specific diagnosis and research in future.
出处
《中国卫生检验杂志》
CAS
2011年第2期386-388,共3页
Chinese Journal of Health Laboratory Technology
基金
海军司令部科研基金资助(08-3309)
关键词
单核李斯特菌
单克隆抗体
ELISA
Listeria monocytogenes
Monoclonal antibody
Sandwich ELISA
