中国媒介生物学及控制杂志 ›› 2019, Vol. 30 ›› Issue (5): 519-523.DOI: 10.11853/j.issn.1003.8280.2019.05.009

• 论著 • 上一篇    下一篇

云南省边境地区卵形硬蜱中中华基因型伯氏疏螺旋体的检出和鉴定

段兴德1, 何志海1, 高子厚1, 蒋宝贵2, 龚正达1, 张云1, 邵宗体1, 江佳富2, 孙毅2, 刘洪波2, 姚明国1, 王帆1, 杜春红1   

  1. 1 云南省地方病防治所医学动物昆虫防制科, 云南 大理 671000;
    2 军事科学院军事医学研究院微生物流行病研究所, 病原微生物生物安全国家重点实验室, 北京 100071
  • 收稿日期:2019-04-18 出版日期:2019-10-20 发布日期:2019-10-20
  • 通讯作者: 杜春红,Email:dch6890728@163.com
  • 作者简介:段兴德,男,彝族,副主任医师,从事自然疫源性疾病防治研究,Email:402284907@qq.com
  • 基金资助:
    国家自然科学基金(81760607,81360413);云南省技术创新人才培养项目(2014HB093);病原微生物生物安全国家重点实验室开放基金(SKLPBS1833)

Detection and identification of Borrelia sinica in Ixodes ovatus from the border region of Yunnan province, China

DUAN Xing-de1, HE Zhi-hai1, GAO Zi-hou1, JIANG Bao-gui2, GONG Zheng-da1, ZHANG Yun1, SHAO Zong-ti1, JIANG Jia-fu2, SUN Yi2, LIU Hong-bo2, YAO Ming-guo1, WANG Fan1, DU Chun-hong1   

  1. 1 Yunnan Institute for Endemic Diseases Control and Prevention, Dali 671000, Yunnan Province, China;
    2 State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences
  • Received:2019-04-18 Online:2019-10-20 Published:2019-10-20
  • Supported by:
    Supported by the National Natural Science Foundation of China (No. 81760607, 81360413), Yunnan Provincial Training of Youth Leader in Academic and Technical Reserve Talents (No. 2014HB093), and the State Key Laboratory of Pathogen and Biosecurity of China (No. SKLPBS1833)

摘要: 目的 对云南省耿马傣族佤族自治县(耿马县)采集的蜱进行伯氏疏螺旋体分子生物学检测,为当地莱姆病的调查提供参考。方法 对2016年4月在云南省耿马县采集的游离蜱单只进行DNA提取后,采用巢式PCR扩增伯氏疏螺旋体5S~23S rRNA间隔区作为初筛实验,阳性者进一步检测16S rRNAFLA鞭毛蛋白基因确证。结果 3种蜱共94只,检出阳性14只,阳性率为14.89%。阳性蜱均为卵形硬蜱,不同性别间差异无统计学意义(χ2=0.746,P=0.388)。锐跗硬蜱和金泽革蜱均未检出阳性。检出的伯氏疏螺旋体3个目的基因片段序列均与中华基因型伯氏疏螺旋体同源性达98%~99%,构建的系统发育树显示,与我国四川、安徽省检出的中华基因型伯氏疏螺旋体聚在一个分支上;与日本疏螺旋体接近,而阿弗西尼疏螺旋体、狭义伯氏疏螺旋体、法雷斯疏螺旋体等基因型聚在外围。结论 耿马县采集的卵形硬蜱携带中华基因型伯氏疏螺旋体,其宿主和媒介的种类、分布及其对人的致病性等,值得进一步调查研究。

关键词: 卵形硬蜱, 伯氏疏螺旋体, 中华基因型伯氏疏螺旋体

Abstract: Objective To identify Borrelia species in ticks collected in Gengma Dai and Va Autonomous county of Yunnan province using molecular biology detection methods, and to provide a basis for the investigation of local Lyme disease. Methods DNA was extracted from individual free-living ticks collected in Gengma county of Yunnan province in April 2016, and Borrelia DNA was identified by nested-PCR amplification of the 5S-23S rRNA intergenic spacer region. Positive samples were further tested by 16S rRNA amplification and verified with the flagellin (FLA) gene. Results A total of 94 ticks belonging to 3 species were captured, and 14 ticks (14.89%) were positive for Borrelia. All Borrelia-positive ticks were Ixodes ovatus, and there was no significant difference in the rate of Borrelia positivity between sexes (χ2=0.746, P=0.388). Ixodes acutitarsus and Dermacentor auratus tested negative for Borrelia. The sequences of the three target gene fragments identified from Borrelia were 98%-99% homologous to those in B. sinica. Phylogenetic analysis showed that the bacterial species identified in this study clustered with B. sinica detected in Sichuan and Anhui provinces, China, and was close to B. japonica but distinct from B. afzelii, B. burgdorferi sensu stricto, and B. valaisiana. Conclusion Ixodes ovatus collected from Gengma county carries B. sinica. The types and distribution of hosts and vectors of B. sinica and its pathogenicity in humans warrant further investigation.

Key words: Ixodes ovatus, Borrelia burgdorferi, Borrelia sinica

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