本文關(guān)鍵詞： 沙眼衣原體 Pgp3 OmcB CT841 Hsp-60　出處：《天津醫(yī)科大學》2014年碩士論文　論文類型：學位論文

【摘要】：沙眼衣原體(Chlamydia trachomatis,C.t)引起的泌尿生殖道感染是近年來國內(nèi)外最常見的性傳播疾病,目前全球范圍每年近10億人被臨床診斷為衣原體感染。多數(shù)衣原體生殖道感染初期癥狀輕微、病情隱匿、難于被發(fā)現(xiàn),感染常常持續(xù)存在,最終導致嚴重的并發(fā)癥,如女性不孕、異位妊娠,男性前列腺炎、附睪炎等。大約70%-90%的女性感染者和50%-70%的男性感染者是沒有癥狀的,他們長期攜帶衣原體,成為重要儲主和傳染源。這就解釋了泌尿生殖道沙眼衣原體感染在全世界持續(xù)流行的根源。 為控制沙眼衣原體感染,早期發(fā)現(xiàn)沙眼衣原體感染者并給與治療,防止進一步傳播是重要舉措之一。但是,目前我國使用的衣原體檢測試劑并不適合早期篩查�，F(xiàn)有檢測試劑的抗原蛋白是衣原體主要外膜蛋白(major outer membrane protein,MOMP)或者熱休克蛋白60(chlamydial heat shock protein60,Hsp-60),近幾年的研究顯示,這兩種蛋白的抗原性都不夠強。 本實驗選取了幾種近年來國外陸續(xù)報道的沙眼衣原體免疫優(yōu)勢蛋白進行蛋白的表達、驗證,以Hsp-60為對照,收集臨床沙眼衣原體感染者和未感染者的血清,ELISA方法檢測血清中抗所表達衣原體抗原的抗體,尋找適用于國內(nèi)早期篩查泌尿生殖道沙眼衣原體感染者的抗原蛋白。 目的：表達三種沙眼衣原體免疫優(yōu)勢蛋白,以熱休克蛋白60為對照,驗證表達蛋白的抗原性,探索適合于早期篩查衣原體感染者的抗原物質(zhì)。 方法：將編碼衣原體質(zhì)粒編碼蛋白3(Pgp3)、外膜蛋白復(fù)合物B蛋白C端肽(OmcBc)、ct841編碼蛋白(CT841)和熱休克蛋白60(Hsp-60)四種基因的重組質(zhì)粒和空質(zhì)粒轉(zhuǎn)化入大腸桿菌,IPTG誘導表達,并用谷胱甘肽磁珠進行純化后包被ELISA孔板。收集天津醫(yī)科大學總醫(yī)院性病門診泌尿生殖道沙眼衣原體感染者和未感染者的血清各20份。ELISA方法檢測血清中上述幾種蛋白的抗體。 結(jié)果：20份沙眼衣原體感染者血清中Pgp3、OmcBc、CT841和Hsp-60抗體的檢出率分別是14例(70%)、9例(45%)、8例(40%)、5例(25%)。20份無沙眼衣原體感染者的血清中,1例只檢測到Hsp-60抗體,其余均無上述4種蛋白抗體的存在。 結(jié)論：上述四種衣原體蛋白中,Pgp3的抗原性最強,抗體檢出率最高；Hsp-60的抗原性最弱,抗體檢出率最低。
[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.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R691.3

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本文編號：1494481