本文選題：結(jié)核分枝桿菌 + 抗體�。� 參考：《延邊大學(xué)》2009年碩士論文

【摘要】： [目的]分離純化五種結(jié)核分枝桿菌分泌性蛋白(30-kDa,38-kDa,ESAT-6,Rv3872 and HBHA),比較這五種蛋白及三種聯(lián)合蛋白(38-kDa-30-kDa Ag,38-kDa-ESAT-6 Ag,38-kDa-HBHA Ag)的免疫學(xué)特性,并評價(jià)其在結(jié)核病血清學(xué)診斷方面的應(yīng)用價(jià)值。為開發(fā)新型結(jié)核分枝桿菌診斷性試劑,提供理論依據(jù)。 [方法]應(yīng)用硫酸銨鹽析作用、疏水作用和陰離子交換作用從結(jié)核分枝桿菌培養(yǎng)濾過液中分離純化30-kDa和38-kDa蛋白:應(yīng)用金屬螯合NI~(2+)-NTA瓊脂親和層析作用從結(jié)核分枝桿菌培養(yǎng)濾過液中分離純化ESAT-6、Rv3872和HBHA蛋白。在肺結(jié)核患者和正常對照組血清中,利用間接酶聯(lián)免疫吸附試驗(yàn),評價(jià)五種結(jié)核分枝桿菌分泌性蛋白及其三種聯(lián)合蛋白的免疫原性。 [結(jié)果]成功分離純化了5種分泌性蛋白。并且在酶聯(lián)免疫吸附試驗(yàn)中,38-kDa抗原與其他四種結(jié)核分枝桿菌分泌性蛋白比較,具有較強(qiáng)的敏感性和特異性,尤其在抗IgM抗體的誘導(dǎo)免疫反應(yīng)最強(qiáng)(P＜0.0001),其敏感性和特異性分別為44.44%和95%。而在38-kDa與30-kDa聯(lián)合抗原中敏感性高達(dá)80%,并且也具有較高的特異性。結(jié)果還顯示出,ESAT-6抗原和肝素結(jié)合血凝黏附素(HBHA)抗原,在與三種抗體的反應(yīng)中,具有相似的敏感性與特異性。Rv3872抗原比其他四種抗原的敏感性和特異性均低。 [結(jié)論]1.作為結(jié)核分枝桿菌分泌性蛋白單一抗原,38-kDa抗原的免疫原性優(yōu)于其他四種抗原(30-kDa,ESAT-6,Rv3872 and HBHA)。2.38-kDa與30-kDa聯(lián)合抗原,有希望成為結(jié)核病血清學(xué)新型診斷試劑有效的候選抗原。
[Abstract]:Separation and purification of five secreted proteins of Mycobacterium tuberculosis to (30-kDa, 38-kDa, ESAT-6, Rv3872, and, HBHA) between the five proteins and three combined protein (38-kDa-30-kDa Ag, 38-kDa-ESAT-6 Ag, 38-kDa-HBHA Ag) on the immunological characteristics, and to evaluate its application value in serological diagnosis of tuberculosis. For the development of new Mycobacterium tuberculosis diagnostic reagents, and provide a theoretical basis.
[method] the application of ammonium sulfate precipitation, hydrophobic interaction and anion exchange separation and purification of 30-kDa and 38-kDa culture filtrate protein from Mycobacterium tuberculosis: application of metal chelating NI~ (2+) -NTA agar affinity chromatography purification effect of ESAT-6 filtration liquid from Mycobacterium tuberculosis culture, Rv3872 and HBHA protein in patients with pulmonary tuberculosis. And the normal control group in serum by indirect enzyme linked immunosorbent assay, the immunogenicity of secretory protein and its three joint evaluation of five protein of Mycobacterium tuberculosis.
[results] the successful purification of 5 kinds of secretory proteins. And in ELISA, 38-kDa antigen and other four secreted proteins from Mycobacterium tuberculosis, and has high sensitivity and specificity, especially in the induction of immune response to the strongest anti IgM antibody (P < 0.0001), its sensitivity and specificity of 44.44% and 95%. respectively in 38-kDa and 30-kDa and the sensitivity of combined antigen in up to 80%, and also has a high specificity. The results also showed that ESAT-6 antigen and heparin binding hemagglutinin adhesin antigen (HBHA), and in three kinds of antibody response, with similar sensitivity and specificity of.Rv3872 antigen than the other four antigens of sensitivity and specificity are low.
[conclusion]1. as a single antigen of Mycobacterium tuberculosis secretory protein, the immunogenicity of 38-kDa antigen is superior to the other four antigens (30-kDa, ESAT-6, Rv3872 and HBHA),.2.38-kDa and 30-kDa combined antigen. It is hoped to become an effective candidate antigen for new diagnostic reagent of tuberculosis serology.

【學(xué)位授予單位】：延邊大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R392

【參考文獻(xiàn)】

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1 全國結(jié)核病流行病學(xué)抽樣調(diào)查技術(shù)指導(dǎo)組;2000年全國結(jié)核病流行病學(xué)抽樣調(diào)查報(bào)告[J];中國防癆雜志;2002年02期

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