文章摘要
刘英,黄荷,张翠彩,姚光海,徐飞,马青,王月,唐光鹏,王定明,李世军.贵州省不明原因发热病例钩端螺旋体病原体分离鉴定研究[J].中华流行病学杂志,2020,41(7):1110-1114
贵州省不明原因发热病例钩端螺旋体病原体分离鉴定研究
Isolation and identification of Leptospira in patients with fever of unknown origin in Guizhou province
收稿日期:2019-10-14  出版日期:2020-07-15
DOI:10.3760/cma.j.cn112338-20191014-00733
中文关键词: 钩端螺旋体;发热病例;聚合酶链式反应;多位点序列分型
英文关键词: Leptospira;Patients with fever symptom;PCR;Multiple locus sequence typing
基金项目:国家自然科学基金(81960622,81760366,81601812);贵州省重要传染病实验诊断技术及分子流行病学研究科技创新人才团队专项资金[黔科合平台人才(2018)5606];贵州省高层次创新型人才培养[黔科合人才(2016)4021];贵州省疾病预防控制中心基金(2017-E2-1)
作者单位E-mail
刘英 贵州省疾病预防控制中心, 贵阳 550004  
黄荷 贵州省疾病预防控制中心, 贵阳 550004  
张翠彩 中国疾病预防控制中心传染病预防控制所, 北京 102206  
姚光海 贵州省疾病预防控制中心, 贵阳 550004  
徐飞 贵州省疾病预防控制中心, 贵阳 550004  
马青 贵州省疾病预防控制中心, 贵阳 550004  
王月 贵州省疾病预防控制中心, 贵阳 550004  
唐光鹏 贵州省疾病预防控制中心, 贵阳 550004  
王定明 贵州省疾病预防控制中心, 贵阳 550004  
李世军 贵州省疾病预防控制中心, 贵阳 550004 zjumedjun@163.com 
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中文摘要:
      目的 对贵州省钩端螺旋体(钩体)病疫区不明原因发热病例进行钩体分离鉴定和分子分型,了解其病原学特征,为当地钩体病的防治提供病原学依据。方法 采集贵州省钩体病疫区黔东南州黎平县不明原因发热病例血液和尿液标本进行钩体分离培养,对分离的钩体疑似菌株通过致病性钩体G1/G2-PCR方法进行初步鉴定,进一步采用钩体血清群特异PCR进行分群鉴定,然后采用多位点序列分型(MLST)技术对其进行分子分型,并与国内常见血清群参考菌株进行聚类分析。结果 从35例发热病例血液中分离得到3株钩体疑似菌株,分离率为8.6%,分别命名为17BX002、17BX003和17AJX008;3株菌株经钩体特异性G1/G2-PCR鉴定为致病性钩体;钩体血清群PCR鉴定显示,17BX002株为流感伤寒群钩体,其余2株菌为阴性(排除其为黄疸出血群、赛罗群、犬型、秋季群、流感伤寒群和七日热群);进一步的MLST显示,17BX002株为ST106型,与流感伤寒群聚类最近,而其余2株为ST96型,与巴达维亚群菌株一致。结论 高发季节贵州省疫区不明原因发热病例中存在钩体感染病例,流感伤寒群和巴达维亚群为贵州省新发现钩体菌群。
英文摘要:
      Objective To understand the etiological characteristics of the patients with fever of unknown origin in Guizhou province through the isolation and identification of Leptospira interrogans and provide evidence for the control, prevention and treatment of human leptospirosis. Methods Blood and urine samples were collected from patients with fever symptoms in Qiandongnan, an epidemic area, in Guizhou. The suspected Leptospira strains were primarily identified using pathogenic Leptospira specific G1/G2-PCR, and subsequently identified by using Leptospira serogroups specific PCR. The Leptospira strains were then genotyped with multiple locus sequence typing. MLST data based cluster analysis on the isolates and Leptospira reference strains of common serogroups were analyzed by using software NTsys 2.10e. Results Three suspected strains of Leptospira were isolated from human blood samples, the isolation rate was 8.6%, which were designated as strain 17BX002, 17BX003 and 17AJX008. Strain 17BX002 was further identified as serogroup grippotyphosa by using Leptospira serogroup specific PCR, while the other two strains were negative (excluded as iterohaemorrhagiae, sejroe, canicola, autumnalis, grippotyphosa and hebdomadis). MLST genotyping showed that strain 17BX002 was typed as ST106, most closely clustered with Leptospira grippotyphosa, while strain 17BX003 and 17AJX008 were typed as ST96, the same as serogroup badaviae. Conclusion There are leptospirosis cases in epidemic area of Guizhou in high incidence season, grippotyphosa and bataviae are the newly discovered serogroups of Leptospira in Guizhou.
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