黑龙江尚志地区莱姆病伯氏疏螺旋体的鉴定与遗传进化分析  

Identification and phylogenetic analysis of Lyme disease Borrelia spp. isolated from Shangzhi Prefecture of Heilongjiang Province

作　　者：牛庆丽[1] 杨吉飞[1] 关贵全[1] 付钰广[1] 马米玲[1] 李有全[1] 刘军龙[1] 刘志杰[1] 刘爱红[1] 任巧云[1] 郝雪峰[1] 罗建勋[1] 殷宏[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部草食家畜疫病重点实验室甘肃省动物寄生虫病重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2010年第7期661-666,共6页Chinese Veterinary Science

基　　金：国家科技支撑计划项目(2007BAD40B02);国家自然资源平台项目(2005DKA21100);国家自然科学基金项目(30571397);国家高技术研究发展计划(863)项目(2006AA10A207);科技部欧盟项目专项经费;欧盟EPI-ZONE;ICTTD3(INCONo;510561)

摘　　要：自我国黑龙江省尚志市野外采集的蜱体内分离出1株莱姆病伯氏疏螺旋体,并对其进行了形态观察及分子生物学分析。利用BSK H分离培养法,经暗视野显微镜和姬姆萨染色观察,菌体呈螺旋状。根据已报道的伯氏疏螺旋体16S rRNA基因序列设计引物,结合参考文献设计的外膜蛋白A(OspA)基因引物经PCR扩增并测序,与GenBank中的各种伯氏疏螺旋体基因型建立系统发生树并进行同源性分析。结果表明,16S rRNA基因核苷酸序列与GenBank中的伽氏疏螺旋体(Borrelia garinii)DK27(X85193)具有较高的同源性,可达99.5%;OspA基因核苷酸序列与报道的Borrelia gariniiKhab(AY502600)有较高的同源性,为93.6%。根据序列分析结果,认为该分离株属于Borrelia garinii基因型,命名为SZ,该菌株16SrRNA基因和OspA基因核酸序列已提交到GenBank(登录号分别为HM007279和HM007278)。A strain of Lyme disease Borrelia burgdorferi was isolated from field ticks collected in Shangzhi Prefecture of Heilongjiang Province,and its morphology and molecular characteristics were studied.Helical structure of the cultured bacterium in BSK H medium was observed on Giemsa stained slides under microscope with dark field.Genomic DNA of the strain was amplified by 16S rRNA and OspA gene based PCR,and the PCR products were sequenced.The sequences of 16S rRNA and OspA gene were compared to that of other Borrelia burgdorferi strains available in GenBank.Two phylogenetic trees were constructed.The results revealed that the 16S rRNA gene sequence shared 99.5% similarity to that of Borrelia garinii strain DK27（X85193）,and the OspA gene sequence shared 93.6% similarity to that of B.garinii strain Khab（AY502600）.According to the sequences analysis,the strain could be identified to be B.garinii,and named SZ.The sequences have been submitted to GenBank（Accession No.：HM007279,HM007278）.

关 键 词：伯氏疏螺旋体 分离 鉴定 系统发生树 

分 类 号：S852.63[农业科学—基础兽医学]