16S rRNA序列同源性分析结合系统发育树构建鉴定6株生殖道乳杆菌  

16S rDNA Identification and Phylogenetic Analysis of Six Strains of Lactobacillus from Female Genital Tract

作　　者：何亮[1] 谢小军 王冲 曾忠铭[1] 

机构地区：[1]广东医学院附属南山医院检验科,广东省深圳市518052 [2]广东省深圳市西丽医院检验科,518000 [3]广东省深圳市南山妇幼保健院检验科,518000

出　　处：《中华全科医学》2013年第4期617-618,640,F0003,共4页Chinese Journal of General Practice

基　　金：广东省深圳市科技计划项目(201103310)

摘　　要：目的对6株分离自健康妇女生殖道乳杆菌进行16S rDNA鉴定和系统发育分析。方法采用Nugent评分进行镜检,筛选健康女性阴道分泌物进行乳杆菌纯培养,抽提染色体DNA,16S rRNA PCR扩增,构建系统发育树。结果采用16S rRNA序列同源性分析方法与系统发育树构建,鉴定201101菌株、201102菌株、201103菌株、201201菌株和201202菌株分别相应与唾液乳杆菌、加氏乳杆菌、詹氏乳杆菌及卷曲乳杆菌形成一个分支,验证可信度均达到100%;201104菌株16S rDNA序列与鸡乳杆菌的16S rDNA序列同源性达99%,系统发育树验证可信度达到98%以上。结论 16S rRNA序列同源性分析与系统发育树构建相结合,可以较准确鉴定细菌种类。Objective For being further applied and studied,six strains of Lactobacillus separated from female genictal tract were identified by 16S rDNA and phylogenetic analysis.Methods Nugent standard was used to diagnose the vaginal discharge.MRS agar was used to culture Lactobacillus.The 16S rDNA of Lactobacillus was amplified with the genomic DNA of six strains of Lactobacillus from female genital tract.Nucleic acid sequence was blastn in GenBank and phylogenetic tree was constructed using neighbor-joining method of distance nethods.Results On the phylogenetic tree,201101 strain formed a separate branch with Lactobacillus salivarius strain.201102 strain formed a separate branch with Lactobacillus gasseri strain.201103 strain formed a separate branch with Lactobacillus jensenii strain.201201 strain and 201202 strain formed a separate branch with Lactobacillus crispatus strain.Bootstrap values were all 100%.The homology of 16S rDNA of 201104 strain with the 16S rDNA of Lactobacillus gallinarum strain was 99%,and Bootstrap value was 98%.Conclusion Combining 16s rRNA sequence homology analysis with phylogenetic tree construction can more accurate identification of bacteria species.

关 键 词：16SrDNA 系统发育树 乳杆菌 生殖道 

分 类 号：R446.51[医药卫生—诊断学] R446.6[医药卫生—临床医学]