迟缓爱德华氏菌EcoRⅠ核糖体自动化分型与聚类分析

Characterization of Edwardsiella tarda by automated EcoRⅠ ribotyping and cluster analysis

摘要:

【目的】对26株迟缓爱德华氏菌进行自动化核糖体分型，并进行聚类分析。【方法】采用RiboprinterTM全自动微生物鉴定系统对分离自斑点叉尾鮰、日本鳗、多宝鱼、比目鱼、斑醴等宿主体内的迟缓爱德华氏菌进行核糖体分型，以限制性内切酶EcoRⅠ处理、切割菌株DNA；运用BioNumerics软件分析图像数据。【结果】迟缓爱德华氏菌核糖体图谱与数据库中已有信息进行比对，ATCC15947和BYK00685的比对相似值>0.85，分别为0.95及0.90。条形码经软件分析共产生21种核糖体条带，聚类分析分为3个群。条带之间呈现明显的地域性差异和宿主差别。来自北方及南方的菌株除少数几株以外，各自聚集为一个群；所有人源株则分布在第三群。【结论】自动化核糖体分型可以方便快捷地用于不同物种的菌株分型与流行病学追踪溯源。

Abstract:

[Objective] A total of 26 Edwardsiella tarda isolates were characterized by automated ribotyping and cluster analysis. [Methods] Strains of E. tarda taken from Channel Catfish, Japanese eel, Turbot, flounder, sweet spot were analyzed by the DuPont RiboprinterTM Microbial Characterization System and we adopted a protocol using EcoRⅠ as restriction enzyme to differentiate and characterize the isolates discussed here. The Riboprinter patterns obtained for the 26 isolates were imported and analyzed with BioNumerics software. [Results] The ribotype profiles of the isolates were compared with the reference DuPont identification database. Only 2 isolates of ATCC15947 and BYK00685 matched the patterns of DuPont Identification Library with a similarily >0.85, reached 0.95 and 0.90. Altogether, 21 different ribotypes (RTs) were generated. There were geographical differences between the RTs, E. tarda isolates collected from host in Northern China were phylogenetically from isolates from Southern China were clustered into two distinct clades, all isolates collected from human and other fishes belonged to third clade. [Conclusion] The discrimination power of automated ribotyping can be used to characterize bacterial isolates of various speices for epidemiological studies.