中国人兽共患病学报2011，Vol.27Issue(7)：587-591,5.
贵州省3株钩端螺旋体分离株PFGE分型与基因种鉴定
Leptospira interrogans isolates by pulsed-field gel electrophoresis typing and genospecies identification in Guizhou Province
摘要
Abstract
The aim of the study was to perform a molecular epidemiological investigation on the Leptospira isolates from animal hosts in Guizhou province by using pulsed-field gel etectrophoresis (PFGE) molecular typing and 16S rRNA gene sequencing based species-specific identification. The extracted chromosomal DNA from leptospiral isolates were digested with restriction endonuclease NotⅠand the DNA segments were separated by using PFGE. By BioNumerics V4.0 software and 75％ similarity as the standard, the obtained PFGE images from leptospiral isolates were managed and then the PFGE maps of leptospiral isolates were compared with those of reference strains belonging to 15 Serovar 15 serogroups of L. interrogans for cluster analysis. Besides, species-specific identification of those three Leptospira isolates were performed by 16S rRNA gene sequencing and analysis. PFGE analysis indicated that 3 strains of L. interrogans from Guizhou province were classified in to two PFGE types, and both of the two PFGE types belonged to the Serovar-groups Icterohaemorrhagiae Serovar Lai strain Lai. 16S rRNA gene sequencing and analysis demonstrated the sequences of 16S rRNA gene of the 3 strains were identical, with homogeny as 100％. And the homogeny of 16S rRNA gene between the three isolated strains and Leptospira reference strains of different serovar types contained in L. interrogans species was 100％. The results revealed that 3 strains of L. interrogans isolated from Guizhou province belonged to Serovar-groups Icterohaemorrhagiae Serovar Lai, and belonged to L. interrogans genospecies. The identification results of PFGE typing are consistent with that of 16S rRNA analysis. It would contribute to the active surveillance, investigation of outbreak and source tracking for leptospirosis in Guizhou province.关键词
钩端螺旋体/PFGE/16S rRNA/序列分析/同源性/基因种Key words
L. interrogans/ PFGE/ 16S rRNA/ sequence analysis/ homogeneity/ genospecies分类
医药卫生引用本文复制引用
李世军,蒋秀高,张翠彩,李秀文,唐光鹏,田克成,刘英,蔡星和,于春,聂一新..贵州省3株钩端螺旋体分离株PFGE分型与基因种鉴定[J].中国人兽共患病学报,2011,27(7):587-591,5.基金项目
贵州省优秀科技教育人才省长专项资金项目(No.黔省专合字(2010)90号)与贵州省疾病预防控制中心青年科学基金项目(No.2009-(青2)-2)联合资助(李世军与张翠彩同等贡献) （No.黔省专合字(2010）
