【摘要】：目的 應用PFGE技術對我國布魯氏菌的不同年代、不同種型菌株和我國分離非典型菌株多態(tài)性進行分析，建立適合布魯氏菌的核算內(nèi)切酶及實驗方法，探索分型研究。將布氏菌的PFGE資料輸入PulseNeL China網(wǎng)絡數(shù)據(jù)庫，作為中國布氏菌的基本數(shù)據(jù)。為布氏菌分了生物學的網(wǎng)絡研究奠定基礎。為我國布魯氏菌病的分子流行病學研究提供分子生物學研究依據(jù)。 方法 通過對國際參考菌株的實驗建立了適合于布氏菌的PFGE實驗方法。進一步選擇確定了核酸內(nèi)切酶XbaI。通過多次對參考菌株的實驗，驗證了實驗的重復性，，穩(wěn)定了實驗條件。進一步對122株國內(nèi)分離菌株進行PFGE檢測。 結果 通過參考菌株的XbaI PFGE圖譜分析，牛種、羊種菌和豬1、3型是穩(wěn)定的型別，而其余的則表現(xiàn)為相似度較差，這一點符合布氏菌的分類歷史，即，犬種菌是牛種菌的轉移宿主形成；其他菌株是野生動物體內(nèi)，自然界中循環(huán)、保存的菌株。由此也證實了PFGE方法的客觀性。為布氏菌的分型提供了分子生物學依據(jù)。發(fā)現(xiàn)了不同種或生物型菌株的核酸酶切序列分布特征。 結論 建立了一套布氏菌的PFGE鑒定參考標準分型方法，對布氏菌進行聚類分析和從全基因角度進行分析研究。建立了布氏菌的PulseNet China網(wǎng)絡數(shù)據(jù)庫，適于常見、對人感染意義較大的羊、牛、豬種和部分犬種布氏菌分型研究，所輸入的布氏菌PFGE資料已作為中國布氏菌的基本數(shù)據(jù)�？梢赃M行國內(nèi)布氏菌的網(wǎng)絡數(shù)據(jù)資料對比研究。開辟了利用網(wǎng)絡研究、鑒定布魯氏菌的新技術平臺。通過對122株國內(nèi)分離菌株PFGE檢測結果，發(fā)現(xiàn)對大部分菌株可以得出較好結果，甚至可解釋新近發(fā)生的布病疫情，應用于我國布魯氏菌的分子流行病學特點分析研究。建立我國的布魯氏菌PulseNet China網(wǎng)絡數(shù)據(jù)席，在我們進一步完善實驗的同時，充實數(shù)據(jù)庫，使其在布氏菌病的研究和防控中有更大的作用。
[Abstract]:Objective to analyze the polymorphism of different strains of Brucella and atypical strains in China by using PFGE technique, and to establish a suitable accounting endonuclease and experimental method for brucellosis, and to explore the typing of Brucella. The PFGE data of brucellosis were input into the PulseNeL China network database as the basic data of brucellosis in China. It lays a foundation for the study of the biological network of Brucella. To provide molecular biological basis for molecular epidemiology of brucellosis in China. Methods A PFGE method suitable for brucellosis was established through the experiment of international reference strains. Further selection confirmed the endonuclease XbaI. The repeatability of the experiment was verified and the experimental conditions were stabilized by several experiments on the reference strains. Further PFGE analysis was carried out on 122 strains isolated in China. Results according to the XbaI PFGE analysis of the strains, the stable type of bovine, sheep and pig strains was found, while the others showed poor similarity, which was consistent with the history of brucellosis. Canine strain is the transfer host of bovine strain. Other strains are circulating and preserved in wild animals and in nature. Therefore, the objectivity of PFGE method is confirmed. It provides molecular biological basis for the typing of brucellosis. The distribution characteristics of nuclease digestion sequences of different strains or biological strains were found. Conclusion A set of PFGE identification standard typing method for brucellosis was established. Cluster analysis and gene analysis of brucellosis were carried out. The PulseNet China network database of brucellosis was established, which is suitable for the typing of brucellosis in sheep, cattle, pig and some canine breeds, which is of great significance to human infection. The data of brucellosis PFGE has been used as the basic data of brucellosis in China. It can be used to compare the data of brucellosis in China. A new technology platform for the identification of brucella by network research has been established. According to the results of PFGE detection of 122 strains isolated in China, it is found that most strains can get better results, and even explain the recent outbreak of brucellosis, which can be applied to the analysis of molecular epidemiological characteristics of Brucella in China. In order to improve the experiment and enrich the database of brucellosis PulseNet China network data sheet in China will play a more important role in the research and control of brucellosis.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R378

【引證文獻】

相關期刊論文 前2條

1 劉國棟;崔步云;劉榮臻;李元凱;;布魯氏菌多重聚合酶鏈反應鑒定研究[J];疾病監(jiān)測;2008年05期

2 周曉艷;陳燕芬;崔步云;陳澤良;姜海;趙鴻雁;樸東日;李蘭玉;王金桃;;我國羊種3型布魯氏菌的多位點序列分型研究[J];中國人獸共患病學報;2011年05期

相關碩士學位論文 前2條

1 周曉艷;中國布氏菌的多位點序列分型研究[D];山西醫(yī)科大學;2011年

2 楊杰;布魯菌MLVA和HOOF分型標準操作方法的建立及應用[D];山西醫(yī)科大學;2012年

本文編號：2387089