肺炎克雷伯菌单基因分子进化分析及多位点序列分型与基因组系统学研究  

Evolution of single-gene molecule of Klebsiella pneumoniae and systematic study on multilocus sequence typing and genome

作　　者：朱健铭 姜如金[1] 吴康乐[1] 张烽[2] 翁幸鐾[3] 孔海深[4] 

机构地区：[1]杭州市余杭区中医院检验科,浙江杭州311106 [2]南通大学附属第三医院检验科,江苏无锡214041 [3]宁波市第一医院检验科,浙江宁波315010 [4]浙江大学医学院附属第一医院传染病诊治国家重点实验室,浙江杭州310009

出　　处：《中华医院感染学杂志》2014年第12期2861-2864,共4页Chinese Journal of Nosocomiology

基　　金：国家"十一五"科技重大专项基金资助项目(2008ZX100-03-004);杭州市科技计划引导基金资助项目(2012027);杭州市卫生科技计划基金资助项目(2010B078);杭州市余杭区重大科技基金资助项目(2012001)

摘　　要：目的比较7株已完成全基因组测序的肺炎克雷伯菌单基因分子进化分析及多位点序列(MLST)分型与基因组系统学研究的结果,研究其亲缘关系。方法采用Roche 454高通量测序技术对1株泛耐药肺炎克雷伯菌JM45株做全基因组测序(完成图),再与NCBI已完成全基因组测序的6株肺炎克雷伯菌9种看家基因进行单个基因分子进化分析,将该9个基因序列串联作MLST分型,并将7株肺炎克雷伯菌的全基因组序列作聚类分析(为Fast Minimum Evolutin法)。结果 JM45株测得一条完整的基因组(染色体)序列及两条质粒序列;因为不同的基因进化率不同,7株肺炎克雷伯菌9种看家基因单个基因序列的分子进化图并不一致;7株肺炎克雷伯菌MLST分型可见JM45株与HS11286株相同(即为同一型),1084株与NTUH-K2044株相同,但7株肺炎克雷伯菌基因组系统学分析可见,JM45株与HS11286株并不相同(即非为同一型),JM45株与HS11286株距离最近,与342株距离最远,其他各株介于二者之间。结论采用单个基因序列的分子进化分析法及MLST分型用作菌株亲缘性关系分析法并不可靠,本研究为国内首个单个看家基因分子进化分析及MLST分型与基因组系统学研究的比较报道。OBJECTIVE To compare the evolutions of single-gene molecule of 7 strains of K lebsiella pneumoniae of which the whole genome sequencing has been completed and study the genetic relationship .METHODS By means of high throughput Roche 454 sequencing technique ,the whole genome sequencing （completed graph） of one strain of pandrug-resistant K .pneumoniae JM45 was performed ,then phylogenetic analysis was performed for 9 kinds of housekeeping genes of six strains of K .pneumoniae ,for which the whole genome sequencing has been completed in NCBI ,the MLST was performed for the nine gene sequences as tandem ,and the phylogenomics （Fast Minimum Evolutin） of the whole genome sequences of 7 strains of K . pneumoniae were observed .RESULTS A complete genome （chromosome ） sequence and 2 plasmid sequences were obtained in JM 45 . Because of the different evolutionary rates ,the molecular evolutionary graphs of the 9 housekeeping genes of the 7 strains of K .pneumon-iae were not the same .MLST of 7 strains of K .pneumoniae showed that JM45 and HS11286 were the same type , strain 1084 was the same as strain NTUH-K2044;but phylogenomics showed that JM45 was different from HS11286 ,and JM45 was the closest to HS11286 ,and the farthest to strain 342 ,and other strains were between the two .CONCLUSIONS It′s not reliable to perform the analysis of molecular evolution of single-gene sequence and MLST for study of genetic relationship .It＇s the first report in China to conduct the systematic study on MLST and genome as well as the evolution analysis of single housekeeping gene .

关 键 词：肺炎克雷伯菌 泛耐药 全基因组测序 多位点序列分型 基因组系统学 

分 类 号：R378.996[医药卫生—病原生物学]