检验医学与临床Issue(23)：3269-3271,3274,4.DOI:10.3969/j.issn.1672-9455.2014.23.015
抗巨细胞病毒pp65抗原单克隆抗体的制备及临床初步比对检测
The preparation of monoclonal antibodies against the CMV pp65 antigen and preliminary clinical comparison test
陈兰兰 1崔京涛 1倪安平 1闫文娟1
作者信息
- 1. 中国医学科学院北京协和医院检验科，北京 100730
- 折叠
摘要
Abstract
Objective To prepare monoclonal antibodies (McAbs) against the CMV pp65 antigen ,and estab‐lish immumofluorescence method for detection of CM V antigenemia in peripheral blood leukocytes .Methods 24 ,48 , 72 h after CMV AD169 infected human embryonic lung fibroblast (MRC‐5) ,CMV antigen was prepared through ul‐trasonic fragmentation and formalin inactivation ,BALB/C mice were immunonized by the CMV pp65 antigens ,and the standard method was used to produce monoclonal antibodies against the CMV pp65 antigen .The prepared mono‐clonal antibodies were used to establish immumofluorescence method for detection of CMV antigenemia in peripheral blood leukocytes ,and the established method was compared to CMV BriteTM kit .Results There was two hybridoma cells producing specific monoclonal antibodies ,and named 13D1‐3 and 29F1‐1 ,respectively .The immunoglobulin sub‐types were IgG1 and IgG2a .After purification ,monoclonal antibodies of ascites fluid also had good immunological characteristics .Compared monoclonal antibody 13D1‐3 with CMV BriteTM kit ,the positive coincidence rate was 88 .8% ,negative coincidence rate was 98 .9% and the total coincidence rate was 97 .3% (kappa= 0 .895) ,predicted high consistency .Conclusion This study might have successfully produced monoclonal antibodies against the CMV pp65 antigen and established immumofluorescence method for detection of CMV antigenemia in peripheral blood leu‐kocytes ,which could indicate high consistency compared with CMV BriteTM kit .关键词
巨细胞病毒/单克隆抗体/抗原血症Key words
Cytomegaovirus/monoclonal antibody/antigenemia引用本文复制引用
陈兰兰,崔京涛,倪安平,闫文娟..抗巨细胞病毒pp65抗原单克隆抗体的制备及临床初步比对检测[J].检验医学与临床,2014,(23):3269-3271,3274,4.
