本文選題：人布魯氏菌病 + PCR��； 參考：《吉林大學(xué)》2009年博士論文

【摘要】： 布魯氏菌病為動物源性疾病,是由細(xì)胞內(nèi)寄生的布魯氏菌引起的傳染-變態(tài)反應(yīng)性人獸共患傳染病,在世界范圍內(nèi)廣泛流行,對畜牧養(yǎng)殖業(yè)的發(fā)展和人類健康造成嚴(yán)重威脅,已成為重要的公共衛(wèi)生問題。布魯氏菌病的診斷方面存在兩個主要的問題:一是快速準(zhǔn)確診斷,二是現(xiàn)有減毒活疫苗的鑒別診斷。 PCR檢測技術(shù)避免操作活菌,并能檢測到標(biāo)本中較低豐度的病原,適合布魯氏菌這種高危險性病原的檢測。用PCR方法對布魯氏菌病早期、復(fù)發(fā)期以及伴隨多種并發(fā)癥的患者及藥物治療后患者進(jìn)行診斷,具有較好的應(yīng)用前景,我國目前尚無應(yīng)用PCR檢測方法在臨床診斷上跟蹤隨訪的研究報道。本研究針對布魯氏菌的保守基因bp26設(shè)計了屬的引物,針對插入性序列IS711設(shè)計了牛、羊、豬種引物,建立了單重和多重PCR檢測方法,在臨床上對免疫學(xué)診斷為患者、可疑患者和非患者人群進(jìn)行檢測,并結(jié)合流行病學(xué)個案數(shù)據(jù)庫資料擬定了進(jìn)一步跟蹤隨訪檢測的對象,為建立人布魯氏菌病標(biāo)準(zhǔn)化PCR診斷方法提供研究依據(jù)。 目前應(yīng)用血清學(xué)方法不能區(qū)分布魯氏菌減毒疫苗株的反應(yīng)與自然感染,發(fā)展基因標(biāo)記疫苗可解決這一問題。M5是我國廣泛應(yīng)用的減毒活疫苗株,本研究應(yīng)用基因同源重組方法,用卡那抗性基因替換部分bp26基因,構(gòu)建了bp26突變株,并根據(jù)布魯氏菌的保守基因建立了能夠區(qū)分bp26突變株與疫苗株的PCR鑒別方法。bp26突變株在胞內(nèi)外的生存能力顯示,bp26突變株在小鼠體內(nèi)的免疫應(yīng)答和免疫保護(hù)效果均低于M5疫苗株,表明bp26基因在M5疫苗株的免疫保護(hù)作用中發(fā)揮了作用�？紤]到bp26基因在免疫原性和保護(hù)性方面的重要性,我們對BP26蛋白進(jìn)行了表達(dá)及純化,并致力于開發(fā)有效的布魯氏菌亞單位疫苗和作為人布魯氏菌病血清學(xué)的特異性診斷抗原用于臨床診斷研究。
[Abstract]:Brucellosis, an animal borne disease, is an infectious and allergic zoonotic disease caused by brucella, which is widely prevalent in the world and poses a serious threat to the development of animal husbandry and human health. Has become an important public health problem. There are two main problems in the diagnosis of brucellosis: one is rapid and accurate diagnosis, the other is the differential diagnosis of the existing attenuated live vaccine. The PCR technique avoids the operation of live bacteria and can detect the pathogens with low abundance in the specimens, which is suitable for the detection of brucellosis, which is a high risk pathogen. PCR was used to diagnose brucellosis in early stage, relapse stage, complicated with many kinds of complications and after drug treatment. It has a good prospect of application. At present, there is no research report on the application of PCR in clinical diagnosis and follow-up. In this study, we designed primers for the conserved gene bp26 of brucella, designed primers for bovine, sheep and pig breeds for inserted sequence IS711, and established single and multiple PCR detection methods. Suspicious patients and non-patients were tested, and the data of epidemiological case database were used to determine the objects for further follow-up detection, which provided the basis for the establishment of standardized PCR diagnosis method for human brucellosis. At present, serological methods can not distinguish the response of attenuated brucellosis vaccine strain from natural infection. The development of gene marker vaccine can solve this problem. M5 is a widely used live attenuated vaccine strain in China. The bp26 mutant was constructed by replacing part of the bp26 gene with the kana-resistant gene. According to the conserved genes of Brucella, a method of PCR identification was established to distinguish bp26 mutant from vaccine strain. The viability of bp26 mutant in vitro and in vitro showed that the immune response and protective effect of Bp26 mutant in mice was lower than that of M5 vaccine strain. The results suggest that bp26 gene plays an important role in the immune protection of M 5 vaccine strain. Considering the importance of bp26 gene in immunogenicity and protection, we expressed and purified BP26 protein, We also focus on the development of effective brucellosis subunit vaccine and clinical diagnosis as a specific diagnostic antigen for human brucellosis.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2009
【分類號】：R516.7;R392

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 劉復(fù)生;;布病的易感人群與預(yù)防[J];中國農(nóng)業(yè)信息;2013年21期

相關(guān)博士學(xué)位論文 前1條

1 孫麗媛;吉林省布魯氏菌流行株種屬鑒定及其耐藥分子機(jī)制的研究[D];吉林大學(xué);2011年

相關(guān)碩士學(xué)位論文 前3條

1 馬琳;人布魯桿菌病的隨訪調(diào)查和診斷方法研究[D];吉林大學(xué);2011年

2 申峰勇;延邊牛布魯氏菌的分離及bp26基因的克隆和原核表達(dá)[D];延邊大學(xué);2012年

3 張劍;牛布魯氏菌毒力基因Virb8間接ELISA方法建立與應(yīng)用[D];山東農(nóng)業(yè)大學(xué);2013年

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本文編號：1964929