云南澜沧巴尔通体的分离鉴定和序列分析  

STUDY ON ISOLATION,IDENTIFICATION AND SEQUENCE ANALYSIS OF BARTONELLA FROM RATTUS BLOOD IN LANCANG OF YUNNAN

作　　者：杨发莲[1] 白鹤鸣[1] 杨慧[1] 于彬彬[1] 

机构地区：[1]云南省地方病防治所,大理671000

出　　处：《现代预防医学》2007年第19期3601-3603,共3页Modern Preventive Medicine

基　　金：国家自然科学基金资助项目(30360093)

摘　　要：[目的]调查澜沧家鼠中巴尔通体的带菌状况,分离培养鉴定并进行序列分析。[方法]将采获的家鼠血接种于含5%去纤维兔血脑心浸液琼脂培基上,置于35℃含5%CO2培养箱中培养分离巴尔通体,同文献[1]报道一致挑选疑似菌落涂片革兰染色镜检进行初筛,镜下有革兰阴性菌的用巴尔通体属特异性引物进行聚合酶链反应(PCR)扩增特异基因片段[枸橼酸合酶基因(gltA)的379bp片段],电泳图中出现目标带即判断为阳性菌株。然后从中随机选出5株作电镜观察并且对其PCR产物测序,将所测核酸序列进行相似性比较及序列分析。[结果]从94份鼠血分离培养出疑似菌落50株,革兰染色镜检均见阴性小杆菌,经PCR证实为阳性37株。电镜下菌细胞呈短杆状、宽0.2～0.3μm、长0.8～1.0μm、菌壁较厚,无鞭毛。5株菌间相似性为93%～95%,其中1株与我国北京从褐家鼠分离的在GenBank中的编号是DQ884394的菌株相似性为99%,其他4株与Heller,R.S.等从以色列褐家鼠中分离的菌株(B.tribocorum)在GenBank中的编号是AJ005494的相似性分别为99%、99%、94%、94%。[结论]澜沧家鼠中存在巴尔通体感染,说明B.tribocorum为该地的主要优势菌。[Objective]To isolate,identify and make sequence analysis of Bartonella strains from rattus in Yunnan in China.[Methods]Blood samples were collected from 94 rattus in Lancang of Yunnan in 2004.All isolates were grown on brain heart infusion agar plates containing 5% defibrinated rabbit blood.The agar plates were incubated at 35℃ in a humidified with 5% CO2 environment for 40 days.All Bartonella-like isolates were examined by Gram staining for primarily screening.The suspected bacteria colonies were carried out PCR amplification with specific Bartonella introducer to amplify specific gene fragment（379 bp of gltA gene）.If the goal band appeared on electrophoresis chart it could be judged that the bacteria were the positive strains.Then 5 Bartonella strains were random selected,and performed thransmission electron micrograph and the PCR products of them were sequenced and analyzed.[Results]The results showed that 50 Bartonella-like strains were isolated from 94 rattus blood samples,and 37 Bartonella strains were identified by PCR.The Bartonella bacteria was small bacillus,with 0.2-0.3 μm wide and 0.8-1.0 μm long.The wall was thicker but with no flagellum under elecronic microscope.The similarities of gltA sequence from 5 Bartonella were from 93% to 95%.One was 99% similar to B.sp.RN52BJ which was isolated from Rattus Norvegicus in Beijing and with the gene serial number of DQ884394 in GenBank.The others were 99%,99%,94% and 94% respectively similar to B.tribocorum which was isolated from Rattus Norvegicus in Israel with the gene serial number of AJ005494 in GenBank.[Conclusion]There is Bartonella infection in rattus in Lancang.B.tribocorum is the preponderant strain in Lancang.

关 键 词：巴尔通体 家鼠 分离鉴定 序列分析 

分 类 号：R378.49[医药卫生—病原生物学]