肠炎沙门菌随机扩增多态性DNA分子分型.docVIP

肠炎沙门菌随机扩增多态性DNA分子分型 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：肠炎沙门菌随机扩增多态性DNA分子分型 1 1 材料与方法 2 12 方法 3 2 结果 4 3 讨论 6 文2：肠炎沙门菌随机扩增多态性DNA分子分型 7 1 材料与方法 8 12 方法 8 2 结果 9 3 讨论 11 参考文摘引言： 12 原创性声明（模板） 13 文章致谢（模板） 14 正文 肠炎沙门菌随机扩增多态性DNA分子分型 文1：肠炎沙门菌随机扩增多态性DNA分子分型 Application of RAPD in molecular typing Salmonella enteritidis isolates Abstract： Objective Molecular typing 12 strai of the Salmonella enteritidis isolated from different The genomic DNA of different Salmonella enteritis strai were extracted and random amplified polymorphic DNA (RAPD) system was developed and random prime were tested to the analysis on 12 strai of Salmonella enteritis isolated from different sources，and the clear and cotant fingerprints were selected for statistical 4 effective prime 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～ RAPD bands were amplified，833 % of them being comparative research of the cluster analysis based on the RAPD bands spectra showed that the similarity coefficient between two different strai varied from 426% to 100%，and 12 Salmonella enteritis strai in this experiment could be classified to 5 groups according to their genetic Using RAPD to subtype and character Salmonella enteritis at molecular level is more rapid and seitive than other typing methods，which indicate broad future of application。 Key words： Salmonella enteritis;RAPD;type 菌株分型对肠炎沙门菌的流行病学调查，以及研究其感染和传播机制具有重要意义。随机扩增多态DNA（Random Amplified Polymorphic DNA， RAPD）具有操作简便、快速、敏感及不需先了解目的基因的核酸序列等优点，目前广泛应用于微生物的分子分型和鉴定、物种亲缘关系的确定及分子流行病学等领域的研究［1-3］。本研究应用RAPD技术对12株不同来源的肠炎沙门菌进行分子分型，分析指纹图谱对其鉴别的价值并确定不同菌株间亲缘关系，为肠炎沙门菌食源性疾病预防控制和鉴别诊断提供新方法。结果报告如下。 1 材料与方法 11 材料 (1)菌株与来源: 12株肠炎沙门菌分别来自病人分离株4株（菌株号为SEWX01～04），生猪肉分离株2株（菌株号为SEWX05，06），熟食样品分离株1株（菌株号为SEYZ08），鸡分离株3株（菌株号为SEYZ11～13），生鸡肉分离株2株（菌株号为SENT19，20）。(2)酶与试剂: Taq DNA 聚合酶， DNA Marker，dNTPs (大连宝生物工程有限公司)；溶菌酶，蛋白酶K，RNA酶，Sarkosyl(上海申能博彩生物工程公司)；其他试剂均为国产分析纯。(3)随机引