肠炎沙门菌随机扩增多态性DNA分子分型
肠炎沙门菌；随机扩增多态性DNA(RAPD)；分型,,肠炎沙门菌；随机扩增多态性DNA(RAPD)；分型,1材料与方法,2结果,3讨论,参考文献
摘要： 目的 对不同来源的12株肠炎沙门菌分离株进行分子分型。方法 分别提取不同菌株基因组DNA，建立和优化随机扩增多态性DNA(RAPD)反应体系,试用22条随机引物对不同菌株进行RAPD扩增，筛选清晰稳定的扩增条带做统计分析。结果 筛选到OPG-03、OPG-06、OPG-07和OPG-13共4条随机引物具有鉴别作用，每一菌株均可扩增出4～10条DNA片段，大多数片段在100～2000bp之间，共扩增出42条RAPD条带，其中共有条带7条，多态性条带35条，多态性为833%。不同来源的肠炎沙门菌分离株之间的遗传相似系数在426%～100%之间，在0850的相似性水平上可将12株肠炎沙门菌分离株分为5个不同的亚型。结论 应用RAPD技术在分子水平上对肠炎沙门菌进行鉴别和分型具有简便、快速和辨别能力高的优势，对肠炎沙门菌的分子流行病学研究具有较好的应用前景。关键词： 肠炎沙门菌；随机扩增多态性DNA(RAPD)；分型
Application of RAPD in molecular typing Salmonella enteritidis isolates
YANG Zhenquan，FANG Weiming，YANG Zhi,et al.
College of Food Science and Engineering(Yangzhou 225001,China)
Abstract： Objective Molecular typing 12 strains of the Salmonella enteritidis isolated from different origins.Methods The genomic DNA of different Salmonella enteritis strains were extracted and random amplified polymorphic DNA (RAPD) system was developed and optimized.22 random primers were tested to the analysis on 12 strains of Salmonella enteritis isolated from different sources,and the clear and constant fingerprints were selected for statistical analysis.Results 4 effective primers named OPG-03,OPG-06,OPG-07 and OPG-13 were selected,with which each strain could be amplified 4 to 10 DNA fragments of 100bp～2000bp.42 RAPD bands were amplified,833 % of them being polymorphic.The comparative research of the cluster analysis based on the RAPD bands spectra showed that the similarity coefficient between two different strains varied from 426% to 100%,and 12 Salmonella enteritis strains in this experiment could be classified to 5 groups according to their genetic relationship.Conclusion Using RAPD to subtype and character Salmonella enteritis at molecular level is more rapid and sensitive than other typing methods,which indicate broad future of application. ......
您现在查看是摘要页，全文长 9643 字符。