中国媒介生物学及控制杂志 ›› 2010, Vol. 21 ›› Issue (2): 131-133.

• 论著 • 上一篇    下一篇

海南省小型兽类巴尔通体的分离培养和序列分析

宋秀平1,刘起勇1,鲁亮1,赵伟2,李贵昌1,栗冬梅1,孙继民3,黄儒婷1   

  1. 1 中国疾病预防控制中心传染病预防控制所媒介生物控制室(北京 102206); 2 海南省疾病预防控制中心; 3 浙江省疾病预防控制中心
  • 收稿日期:2009-10-23 出版日期:2010-04-20 发布日期:2010-04-20
  • 通讯作者: 刘起勇,Email: liuqiyong@icdc.cn
  • 作者简介:宋秀平(1972-),女,主管技师,从事病原生物学方面的研究。
  • 基金资助:

    国家自然科学基金(30371246)

Isolation and sequence analysis of Bartonella in small mammals in Hainan province

SONG Xiu-Ping, LIU Qi-Yong, LU Liang, ZHAO Wei, LI Gui-Chang, LI Dong-Mei, SUN Ji-Min, HUANG Ru-Ting   

  1. 1 National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China; 2 Hainan Center for Disease Control and Prevention; 3 Zhejiang Center for Disease Control and Prevention. *State Key Laboratory for Infectious Disease Prevention and Control, Beijing, China
  • Received:2009-10-23 Online:2010-04-20 Published:2010-04-20
  • Contact: LIU Qi?yong, Email: liuqiyong@icdc.cn
  • Supported by:

    Supported by the National Natural Science Foundation of China (No. 30371246)

摘要:

目的 调查海南省小型兽类巴尔通体感染状况以及携带巴尔通体的种类,为巴尔通体感染的预防控制提供科学依据。方法 用鼠笼法捕获小型兽类,取心脏血,抗凝,取100 μl抗凝血用胰酶大豆肉汤按1∶4稀释后接种于含5%去纤维羊血的胰酶大豆琼脂培养基上,置含5%CO2的培养箱中37 ℃培养45 d。挑选疑似菌落涂片,革兰染色、Gimanez染色后做镜检进行初筛,将革兰阴性杆菌用巴尔通体属特异性引物进行聚合酶链反应(PCR)。然后对其PCR 产物测序,将所测核酸序列提交到GenBank,做相似性比较及序列分析。结果 从65 份样本中分离培养出疑似菌落6株,革兰染色镜检均见阴性小杆菌,Gimanez 染色为红色杆菌, 经PCR 证实6 株均为巴尔通体,其中分离自黄毛鼠和屋顶鼠各2株,针毛鼠和臭鼩鼱各1株。经过序列分析证实,所分离到的巴尔通体与Bartonella rattimassiliensis、 B. tribocorumB. queenslandensis 3种巴尔通体相似度最高。结论 海南省小型兽类中有巴尔通体感染,存在人群感染的风险。

关键词: 巴尔通体, 小型兽类, 序列分析

Abstract:

Objective To determine the current situation of Bartonella infection in and types of Bartonella carrying small mammals in Hainan province, providing evidence for prevention and control of Bartonella infection. Methods The rat?cage method was employed to capture small mammals, of which heart blood was collected and treated with anticoagulants. A 1∶4 dilution of 100 μl anti?coagulation blood and trypsin soy broth was inoculated onto the trypsin soybean agar culture media containing 5% de?fiber sheep blood and cultured in a 5% CO2 incubator at 37 ℃ for 45 d. Suspected colonies were smeared, Gram- and Gimanez?stained and screened under microscopy. The polymerase chain reaction (PCR) was performed for the Gram?negative bacilli with Bartonella genus?specific primers, followed by sequencing of the PCR products. The nucleic acid sequences were submitted to GenBank for similarity comparison and sequence analysis. Results A total of 6 suspected colonies were isolated from 65 samples, all being Gram?negative small bacilli, and red for Gimanez staining. The PCR results confirmed that the six strains were all Bartonella, two isolated from Rattus losea and Rattus brunneusculue each, and one from Niniventer fulvescens and Suncus murinus each. The  sequence  analysis  suggests  that  the  isolated  Bartonella  had the highest similarity with Bartonella  rattimassiliensis, B. tribocorum  and  B. queenslandensis. Conclusion Bartonella  infection  was  found  among  small mammals in Hainan province, indicating risks of infection among human beings.

Key words: Bartonella, Small mammals, Sequencing analysis

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