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采用初染患者血清样本进行肺炎衣原体特异性抗原的基因组筛查。

Genomic screening for Chlamydophila pneumoniae-specific antigens using serum samples from patients with primary infection.

机构信息

Department of Molecular Genetics, Kawasaki Medical School, Kurashiki, Japan.

出版信息

FEMS Microbiol Lett. 2012 Apr;329(2):168-76. doi: 10.1111/j.1574-6968.2012.02520.x. Epub 2012 Feb 24.

Abstract

Chlamydophila pneumoniae, an obligate intracellular human pathogen, causes respiratory tract infections. The most common techniques used for the serological diagnosis of C. pneumoniae infections are microimmunofluorescence tests and commercial serological ELISA tests; these are based on the detection of antibodies against whole chlamydial elementary bodies and lipopolysaccharide/outer membrane protein, respectively. Identification of more specific and highly immunodominant antigens is essential for the development of new serodiagnostic assays. To identify novel specific antigens from C. pneumoniae, we screened 455 genes with unknown function in the genome of C. pneumoniae J138. Extracts of Saccharomyces cerevisiae cells expressing GFP-tagged C. pneumoniae proteins were subjected to Western blot analysis using serum samples from C. pneumoniae-infected patients as the primary antibodies. From this comprehensive analysis, 58 clones expressing C. pneumoniae open reading frames, including hypothetical proteins, were identified as antigens. These results have provided useful information for the development of new serological tools for the diagnosis for C. pneumoniae infections and for the development of vaccines in future.

摘要

肺炎衣原体,一种专性细胞内人类病原体,引起呼吸道感染。用于肺炎衣原体感染血清学诊断的最常用技术是微量免疫荧光试验和商业血清学 ELISA 试验;这些方法分别基于针对整个衣原体原体和脂多糖/外膜蛋白的抗体的检测。鉴定更特异和高度免疫显性的抗原对于开发新的血清诊断检测方法至关重要。为了从肺炎衣原体中鉴定新的特异抗原,我们在肺炎衣原体 J138 的基因组中筛选了 455 个具有未知功能的基因。用感染肺炎衣原体的患者的血清样本作为一抗,对表达 GFP 标签的肺炎衣原体蛋白的酿酒酵母细胞提取物进行 Western blot 分析。通过这项全面分析,鉴定出 58 个表达肺炎衣原体开放阅读框的克隆,包括假定蛋白,这些克隆被鉴定为抗原。这些结果为开发用于肺炎衣原体感染诊断的新血清学工具以及未来疫苗的开发提供了有用的信息。

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