泌尿生殖道沙眼衣原体感染者血清抗衣原体免疫优势蛋白抗体的检测

发布时间：2018-02-07 13:27

本文关键词： 沙眼衣原体 Pgp3 OmcB CT841 Hsp-60　出处：《天津医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：沙眼衣原体(Chlamydia trachomatis,C.t)引起的泌尿生殖道感染是近年来国内外最常见的性传播疾病,目前全球范围每年近10亿人被临床诊断为衣原体感染。多数衣原体生殖道感染初期症状轻微、病情隐匿、难于被发现,感染常常持续存在,最终导致严重的并发症,如女性不孕、异位妊娠,男性前列腺炎、附睾炎等。大约70%-90%的女性感染者和50%-70%的男性感染者是没有症状的,他们长期携带衣原体,成为重要储主和传染源。这就解释了泌尿生殖道沙眼衣原体感染在全世界持续流行的根源。为控制沙眼衣原体感染,早期发现沙眼衣原体感染者并给与治疗,防止进一步传播是重要举措之一。但是,目前我国使用的衣原体检测试剂并不适合早期筛查。现有检测试剂的抗原蛋白是衣原体主要外膜蛋白(major outer membrane protein,MOMP)或者热休克蛋白60(chlamydial heat shock protein60,Hsp-60),近几年的研究显示,这两种蛋白的抗原性都不够强。本实验选取了几种近年来国外陆续报道的沙眼衣原体免疫优势蛋白进行蛋白的表达、验证,以Hsp-60为对照,收集临床沙眼衣原体感染者和未感染者的血清,ELISA方法检测血清中抗所表达衣原体抗原的抗体,寻找适用于国内早期筛查泌尿生殖道沙眼衣原体感染者的抗原蛋白。目的：表达三种沙眼衣原体免疫优势蛋白,以热休克蛋白60为对照,验证表达蛋白的抗原性,探索适合于早期筛查衣原体感染者的抗原物质。方法：将编码衣原体质粒编码蛋白3(Pgp3)、外膜蛋白复合物B蛋白C端肽(OmcBc)、ct841编码蛋白(CT841)和热休克蛋白60(Hsp-60)四种基因的重组质粒和空质粒转化入大肠杆菌,IPTG诱导表达,并用谷胱甘肽磁珠进行纯化后包被ELISA孔板。收集天津医科大学总医院性病门诊泌尿生殖道沙眼衣原体感染者和未感染者的血清各20份。ELISA方法检测血清中上述几种蛋白的抗体。结果：20份沙眼衣原体感染者血清中Pgp3、OmcBc、CT841和Hsp-60抗体的检出率分别是14例(70%)、9例(45%)、8例(40%)、5例(25%)。20份无沙眼衣原体感染者的血清中,1例只检测到Hsp-60抗体,其余均无上述4种蛋白抗体的存在。结论：上述四种衣原体蛋白中,Pgp3的抗原性最强,抗体检出率最高；Hsp-60的抗原性最弱,抗体检出率最低。
[Abstract]:Chlamydia trachomatisC.t) is the most common sexually transmitted disease at home and abroad in recent years. At present, nearly 1 billion people worldwide are clinically diagnosed as chlamydia infection every year. The initial symptoms of most chlamydia genital tract infections are mild. The condition is hidden, difficult to detect, infections often persist, leading to serious complications such as female infertility, ectopic pregnancy, male prostatitis, Epididymitis and so on. About 70-90% of women infected and 50% -70% of men infected with epididymitis are asymptomatic. They carry chlamydia for a long time. This explains the persistent prevalence of chlamydia trachomatis in the world. In order to control Chlamydia trachomatis infection, early detection and treatment of Chlamydia trachomatis infection and prevention of further transmission are important measures. Chlamydia detection reagents used in China are not suitable for early screening. The antigenic proteins used in the present detection reagents are major outer membrane protein mompp or heat shock protein 60 chlamydial heat shock protein 60 Hsp-60. Recent studies have shown that chlamydia major is the main outer membrane protein of Chlamydia monocytogenes (MMP) or heat shock protein 60 chlamydia dial heat shock protein 60 (Hsp-60). The antigenicity of both proteins is not strong enough. In this experiment, several kinds of Chlamydia trachomatis immunoreactive proteins reported abroad in recent years were selected for protein expression, and Hsp-60 was used as control. Serum Elisa was used to detect the antibody against chlamydia trachomatis antigen in clinical chlamydia trachomatis infection and to find the antigenic protein suitable for early screening of chlamydia trachomatis infection in China. Objective: to express three kinds of Chlamydia trachomatis immune dominant proteins and to test the antigenicity of the expressed proteins with heat shock protein 60 (HSP60), and to explore the antigens suitable for early screening of chlamydia infection. Methods: the recombinant plasmids and empty plasmids encoding chlamydia plasmids encoding protein 3Pgp3, outer membrane protein complex B protein C terminal peptide OmcBct841 and heat shock protein 60hsp-60) were transformed into E. coli induced expression by IPTG. Glutathione beads were purified and coated with ELISA pore plate. 20 serum samples of Chlamydia trachomatis infection and 20 uninfected patients were collected from STD clinic of Tianjin Medical University to detect the antibodies against these proteins. Results in 20 sera of chlamydia trachomatis infected patients, Hsp-60 antibodies were detected in 1 out of 20 serum samples of chlamydia trachomatis infected patients. The detection rates of Hsp-60 antibodies were 14 cases and 9 cases, respectively, and 8 cases were compared with that of 5 cases without chlamydia trachomatis infection, and none of the other cases were found to have the four kinds of protein antibodies mentioned above in the sera of 5 patients with chlamydia trachomatis infection and 20 samples of serum samples of chlamydia trachomatis infected patients with Chlamydia trachomatis infection. Conclusion: among the four chlamydia proteins, the antigenicity of PGP 3 is the strongest, the detection rate of antibody is the highest, the antigenicity of Hsp-60 is the weakest, and the detection rate of antibody is the lowest.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R691.3

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