- 作者：Zhou ZHOUAuthor Vitae ; Yi Mou WU ; yimouwu@sina.com ; Li Li CHEN ; Guang Chao LIU ; Liang Zhuan LIU ; An Wen ZHOU ; Jun Hua ZHANG
- 关键词：Chlamydophila pneumoniae ; Major outer membrane protein ; Monoclonal antibody ; ELISA
- 刊名：Biomedical and Environmental Sciences
- 出版年：2012
- 出版时间：December, 2012
- 年：2012
- 卷：25
- 期：6
- 页码：690-696
- 全文大小：296 K
Objective
This paper aims to develop a monoclonal antibodies (MAbs)- based ELISA for detecting Chlamydophila pneumoniae (C. pneumoniae) antigens in humans with the variable domains (VD) 2 and 3 of the major outer membrane protein (MOMPVD2-VD3) and to assess its sensitivity and specificity by comparing with a widely used MAb that is able to recognize the elementary bodies of C. pneumoniae.Methods
MOMPVD2-VD3 were overexpressed in Escherichia coli and purified by affinity chromatography. Mice were immunized with the recombinant antigen, and hybridomas secreting MAbs were screened. Three stable hybridomas clones were selected and named 5D6, 7G3, and 8C9. The MAbs-based ELISA was scrutinized for species-specific recognition with a number of human throat swab samples from Group I (156 patients with typical respiratory illness clinically confirmed before) and Group II (57 healthy donors).
Results
In Group I, 55 positive cases were detected by anti-EB MAb-based ELISA, 51 cases were positive by MAbs 5D6-based ELISA, and 33 and 38 cases were positive by MAb 8C9 and 7G3-based ELISA respectively. Of the 57 samples from Group II ¡°healthy donors¡±, 5 were positive and 52 were negative with both anti-EB and 5D6-based tests, while 2 and 3 positive cases were identified by the other two MAb-based ELISAs respectively.
Conclusion
The novel MOMPVD2-VD3 MAb-based assay may have higher specificity than the anti-EB MAb, which may possibly be used as an alternative tool for the diagnosis of C. pneumoniae infection.