摘要
目的了解贵州省2009-2015年分离自鼠类宿主动物的钩端螺旋体(简称钩体)PFGE带型特征,为当地钩体病的预防和控制提供科学依据。方法对贵州省近年分离自鼠类宿主动物的56株钩体采用致病性钩体特异引物进行PCR扩增鉴定,应用血清分群PCR对其进行血清群鉴定,采用包埋法提取钩体染色体DNA,利用核酸内切酶NotI对DNA进行酶切,酶切片段经PFGE电泳后采用凝胶成像系统拍照获得DNA指纹图谱,采用国家病原体分子分型实验数据分析与传输系统进行处理和聚类分析。结果致病性钩体特异性PCR和血清分群PCR鉴定显示56株钩体均为黄疸出血群钩体株,PFGE分型将其分为38个带型,聚类分析显示多数钩体株与黄疸出血群赖型具有较高的相似度,为主要流行优势带型,少部分钩体株与爪哇群和波摩那群代表菌株具有较高相似性,聚类关系相对较近。结论黄疸出血群赖型是贵州省近年流行的优势型,钩体株PFGE带型具有多样性,可为当地钩体病的监测、暴发调查和传染源追踪提供参考依据。
Objectives To ascertain the molecular epidemiologic characteristics of pathogenic Leptospira isolated from rats in Guizhou Province from 2009 to 2015 and to provide a scientific basis for the control and prevention of leptospirosis in Guizhou Province. Methods Pathogen-specific PCR was used to identify Leptospira isolates, and serogroup-specific PCR was used to identify the serogroup of the isolates. Chromosomal DNA extracted from leptospiral isolates was digested with the restriction endonuclease Not I, and the DNA segments were separated using PFGE. The Chinese National System for Molecular Typing of Pathogens(http://101.200.213.193:8080/pulsenet) was used to manage PFGE images obtained from leptospiral isolates, and the PFGE maps of leptospiral isolates were compared with those of representative strains belonging to L. interrogans for cluster analysis. Results Leptospira-specific PCR identified the 56 Leptospira isolates as pathogenic Leptospira, and serogroup-specific PCR indicated that all 56 isolates belonged to the Icterohaemorrhagiae serogroup of Leptospira. PFGE indicated that the 56 strains of L. interrogans from Guizhou Province could be classified into 38 PFGE types. Cluster analysis indicated that most of the isolates from Guizhou Province were relatively closely clustered with representative strain 56601 of the Icterohaemorrhagiae serogroup. A few isolates were most closely clustered with representative strain 56608 of the Pomona serogroup and a representative strain of the Java serogroup. Conclusion PFGE revealed that the Icterohaemorrhagiae serogroup has been the prevalent serogroup of L. interrogans in Guizhou Province over the past few years. PFGE also revealed the molecular diversity of the isolates in Guizhou Province. These findings should help to control and prevent leptospirosis in Guizhou Province.
作者
刘英
王铭
李世军
姚光海
黄荷
陈旭
王斌
唐光鹏
王定明
LIU Ying;WANG Ming;LI Shi-jun;YAO Guang-hai;HUANG He;CHEN Xu;WANG Bin;TANG Guang-peng;WANG Ding-ming(Guizhou Provincial Center for Disease Control and Prevention,Guiyang 550004,China)
出处
《中国病原生物学杂志》
CSCD
北大核心
2019年第2期186-189,198,共5页
Journal of Pathogen Biology
基金
国家自然科学基金项目(No.81760366)
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项目(No.黔科合平台人才[2018]5606)
贵州省高层次创新型人才培养项目(No.黔科合人才(2016)4021号)
贵州省优秀青年科技人才培养对象专项资金项目(No.黔科合人字(2015)09号)
贵州省卫生计生委科学技术基金项目(No.GZWJKJ2017-1-011)
中国疾病预防控制中心传染病预防控制国家重点实验室开放课题(No.2017SKLID313)
贵州省疾病预防控制中心基金项目(No.2017-E2-1)联合资助
关键词
贵州
钩端螺旋体
PCR
脉冲场凝胶电泳
Guizhou Province
Leptospira
PCR
Pulsed-field gel electrophoresis
