江西省钩端螺旋体分离株脉冲场凝胶电泳分型和分析

目的 采用脉冲场凝胶电泳(PFGE)技术,对江西省钩端螺旋体(钩体)患者及动物宿主中分离的钩体菌株型别进行分子流行病学调查.方法 利用核酸内切酶Not Ⅰ对提取的钩体菌株染色体DNA进行酶切,通过PFGE将DNA片段分离.获得的PFGE图像采用分析软件BioNumerics 4.0进行处理并建立数字化数据库,以相似度＞75%为标准,将获各钩体PFGE图谱与中国15群15型钩体参考标准株进行比较并进行聚类分析.结果 江西省不同地区(的139株钩体可分为46个PFGE型,优势型为LepNot Ⅰ.0071、LepNotⅠ.0072和LepNotⅠ.0043型,分别占所有钩体菌株的28.06%、15.11%和7.19%.139株钩体分离株中,84.89%(118/139)菌株的PFGE图谱与6群6型中国钩体参考标准株基本相符,其中32.37%(45/139)钩体菌株属于黄疸出血群赖型,15.83%(22/139)和15.11%(21/139)钩体菌株分别属于澳洲群澳洲型和爪哇群爪哇型.结论 PFGE是一种快速、准确、高效的钩体分型方法 .黄疸出血群赖型是江西省人群及动物中优势血清型,澳洲群澳洲型和爪哇群爪哇型位于其次.

Molecular typing on Leptospira interrogans isolates from Jiangxi province, by pulsed-field gel electrophoresis

Objective To perform a molecular epideminlogical investigation on the types of Leptospira interrogans isolates from leptospirosis patients and animal hosts in Jiangxi province,using a pulsed-field gel electrophoresis (PFGE).Methods The extracted chromosomal DNA from leptospiral isolates were digested with restriction endonuclease Not Ⅰ and the DNA segments were separated by using PFGE.By BiOnurerics V4.0 software and 75% similarity as the standard,the obtained PFGE images from leptospiral isolates were managed to establish a digitization database and then the PFGE maps of leptospiral isolates were compared with those of reference standard strains belonging to 15 serovars in 15 serogroups of L.interrogans,for cluster analysis.Results 139 strains of L.interrogans isolated from different areas of Jiangxi province were classified into 46 PFGE types.Among the PFGE types,LepNot Ⅰ.0071,LepNotⅠ.0072 and LepNot Ⅰ .0043 were the predominant types that accounting for 28.06%,15.11% and 7.19% of all the leptospiral isolates,respectively.The PFGE maps from 84.89% (118/139) of the 139 leptospiral isolates were found to basically match those of 6 reference standard strains belonging to 6 serovar in 6 serogroups of L.interrogans.In the 118 matched ieptospiral isolates,32.37% (45 strains),15.83% (22 strains) and 15.11% (21 strains)belonged to sero-groups Icterohaemorrhagiae serovar Lai,sero-groups Australis serovar Australis and sero-group Javanica serovar Javanica,respectively.Conclusion PFGE seemed a fast,accurate and effective method for typing of L.interrogans isolates.Serogroup Icterohaemorrhagiae serovar Lai and followed by serogroup Australis serovar Australis as well as serogroup Javanica serovar Javanica were the predominant L.interrogans species in humans and animal hosts in Jiangxi province.