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 16S rRNA and OspA gene based PCR,and the PCR products were sequenced.The sequences of 16S 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 16S 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）.
出处
《中国兽医科学》
CAS
CSCD
北大核心
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)
关键词
伯氏疏螺旋体
分离
鉴定
系统发生树
Borrelia burgdorferi
isolation
idenfication
phylogenetic tree
