布鲁氏菌脉冲场凝胶电泳分型研究
发布时间：2018-12-19 15:15
【摘要】：目的 应用PFGE技术对我国布鲁氏菌的不同年代、不同种型菌株和我国分离非典型菌株多态性进行分析，建立适合布鲁氏菌的核算内切酶及实验方法，探索分型研究。将布氏菌的PFGE资料输入PulseNeL China网络数据库，作为中国布氏菌的基本数据。为布氏菌分了生物学的网络研究奠定基础。为我国布鲁氏菌病的分子流行病学研究提供分子生物学研究依据。 方法 通过对国际参考菌株的实验建立了适合于布氏菌的PFGE实验方法。进一步选择确定了核酸内切酶XbaI。通过多次对参考菌株的实验，验证了实验的重复性，，稳定了实验条件。进一步对122株国内分离菌株进行PFGE检测。 结果 通过参考菌株的XbaI PFGE图谱分析，牛种、羊种菌和猪1、3型是稳定的型别，而其余的则表现为相似度较差，这一点符合布氏菌的分类历史，即，犬种菌是牛种菌的转移宿主形成；其他菌株是野生动物体内，自然界中循环、保存的菌株。由此也证实了PFGE方法的客观性。为布氏菌的分型提供了分子生物学依据。发现了不同种或生物型菌株的核酸酶切序列分布特征。 结论 建立了一套布氏菌的PFGE鉴定参考标准分型方法，对布氏菌进行聚类分析和从全基因角度进行分析研究。建立了布氏菌的PulseNet China网络数据库，适于常见、对人感染意义较大的羊、牛、猪种和部分犬种布氏菌分型研究，所输入的布氏菌PFGE资料已作为中国布氏菌的基本数据。可以进行国内布氏菌的网络数据资料对比研究。开辟了利用网络研究、鉴定布鲁氏菌的新技术平台。通过对122株国内分离菌株PFGE检测结果，发现对大部分菌株可以得出较好结果，甚至可解释新近发生的布病疫情，应用于我国布鲁氏菌的分子流行病学特点分析研究。建立我国的布鲁氏菌PulseNet China网络数据席，在我们进一步完善实验的同时，充实数据库，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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R378
本文编号：2387089
[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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R378
【引证文献】
相关期刊论文 前2条
1 刘国栋;崔步云;刘荣臻;李元凯;;布鲁氏菌多重聚合酶链反应鉴定研究[J];疾病监测;2008年05期
2 周晓艳;陈燕芬;崔步云;陈泽良;姜海;赵鸿雁;朴东日;李兰玉;王金桃;;我国羊种3型布鲁氏菌的多位点序列分型研究[J];中国人兽共患病学报;2011年05期
相关硕士学位论文 前2条
1 周晓艳;中国布氏菌的多位点序列分型研究[D];山西医科大学;2011年
2 杨杰;布鲁菌MLVA和HOOF分型标准操作方法的建立及应用[D];山西医科大学;2012年
本文编号：2387089
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