合肥首例恙虫病病人血中东方体56kDa外膜蛋白部分基因的扩增及进化分析  

PCR AMPLIFICATION AND PHYLOGENETIC ANALYSIS OF THE PARTIAL GENE SEGMENT OF 56 KDA PROTEIN OF ORIENTIA TSUTSUGAMUSHI FROM THE FIRST CASE OF SCRUB TYPHUS IN HEFEI CITY

作　　者：操敏[1] 耿美玲[1] 陈乐如[1] 毛应华[1] 谭伟龙[1] 王长军[1] 朱进[1] 王忠灿[1] 

机构地区：[1]南京军区疾病预防控制中心,南京210002

出　　处：《寄生虫与医学昆虫学报》2013年第1期31-34,共4页Acta Parasitologica et Medica Entomologica Sinica

基　　金：科技部传染病重大专项（201zx10004801旬04_017,2013zxl0004218-008）;全军“十二五’’重大专项（AWS11L009）：全军“十二五”面上项目（CWS11J258）;2010年度南京军区医学科技创新课题资助项目（10blA129）

摘　　要：采用恙虫病胶体金快速诊断试剂对安徽省合肥市一例不明原因发热病人血清进行检测，从该病人血块中提取DNA为模板，用恙虫病东方体特异性引物，PCR扩增出56kDa外膜蛋白部分基因，将扩增目的片段克隆至T载体进行测序并进行基因进化分析。结果显示，该病人血清中抗恙虫病东方体总抗体、抗东方体IgG及抗东方体IgM阳性；患者给予强力霉素治疗后迅速退热。采用PCR从病人血标本中扩增出1320bp东方体56kDa外膜蛋白基因片段，将基因片段测序结果在NCBI上做BLAST比对，显示该基因序列与昆明株和内蒙古株同源基因序列完全一致；进化树分析显示该序列与标准株Kuroki位于同一个分支。研究确认该发热病例为合肥市报告的首例恙虫病，其感染的恙虫病东方体为Kuroki型，推测合肥市可能为恙虫病新疫区。A case of patient with fever for unknown reason in the city of Hefei was diagnosed as Scrub typhus using colloidal gold rapid diagnosis reagent. The partial gene segment of 56-kDa protein was PCR amplified by the O. tsutsugamushi specific primes using DNA template extracted from the blood of the patient. The amplified gene segment was sequenced. Phylogenetic tree was constructed on the gene of 56 kDa protein of 15 strains. It was resulted that all the anti-Or total antibodies, anti-O, tsutsugamushi IgG and anti-O, tsutsugamushi IgM in the serum were positive. The patient ＇ s fever was allayed obviously after doxycycline treatment and was diagnosed as the first case of scrub typhus in the city of Hefei, Anhui province. The gene sequence of 56 kDa protein showed 100% sequence identity with Inner Mongolia and Kunming strains. Phylogenetic analysis showed that Hefei strain was in the same branch with Inner Mongolia and Kunming strain, and the three were also in the same branch with Kuroki. This study indicated that the city of Hefei in Anhui province may be a new endemic foci of scrub typhus and the genotype of Hefei was determined as Kuroki type.

关 键 词：恙虫病东方体 56 kDa外膜蛋白 序列分析 进化树 

分 类 号：Q987[生物学—遗传学]