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[通过抗菌药物耐药模型、质粒图谱分析和脉冲场凝胶电泳对在土耳其安卡拉分离的志贺氏菌菌株进行特征分析]

[Characterization of Shigella strains isolated in ankara, Turkey by antimicrobial resistance models, plasmid profile analysis and pulsed-field gel electrophoresis].

作者信息

Saran Begüm, Erdem Birsel, Tekeli Fazıl Alper, Sahin Fikret, Aysev Ahmet Derya

机构信息

Ankara University Faculty of Medicine, Ibn-i Sina Hospital, Department of Medical Microbiology, Ankara, Turkey.

出版信息

Mikrobiyol Bul. 2013 Jan;47(1):35-48. doi: 10.5578/mb.4438.

Abstract

Shigella is one of the most important causative agents of diarrhea especially in childhood. Since man is the main reservoir of Shigella and human to human transmission is possible, Shigella can easily spread in public and cause outbreaks. In this study, a total of 60 Shigella strains isolated in Ankara, Turkey by years 2001, 2008 and 2009 were investigated by their antimicrobial susceptibility profiles, plasmid profile analysis (PPA) and pulsed-field gel electrophoresis (PFGE). For epidemiological investigation, the results obtained by antibiotic resistance typing (ART) which was the phenotyping method, was compared to the results of the genotyping methods which were PPA and PFGE. Of the isolates 49 (81.6%) were S.sonnei, 10 (16.6%) were S.flexneri and one was (1.6%) S.dysenteriae. Antimicrobial susceptibilities were evaluated by disc diffusion method and the highest resistance rates were found against trimethoprim/sulfamethoxazole (91.6%), followed by tetracycline (68.3%) and ampicillin (26.6%). Resistance against ampicillin, chloramphenicol and amoxycillin/clavulanic acid were found higher in S.flexneri isolates than S.sonnei (p< 0.001). All isolates were found to be susceptible to ciprofloxacin, gentamicin and ceftazidime. S.sonnei demonstrated 12 and S.flexneri demonstrated 4 antibiotic resistance models. All isolates were carrying plasmids with varying sizes and varying numbers between 1 to 7. S.sonnei isolates demonstrated 27 and S.flexneri isolates demonstrated 8 plasmid profiles. S.sonnei isolates were clustered in 4 patterns and S.flexneri were clustered in 5 patterns by PFGE. This method demonstrated obvious clonal similarity among S.sonnei strains isolated in Ankara and discriminative power (DP) was calculated as 0.26. PPA and ART demonstrated higher DP among S.sonnei strains (0.97 and 0.75, respectively). In this study gain or loss of instable genetic mobile elements were thought to be responsible for higher discriminative powers of PPA and ART methods. These typing methods were found to be appropriate for the epidemiological investigation of strains collected in a short time period. PFGE was found to be convenient for the evaluation of clonal relatedness of the strains, however, in such geographical areas where the same clone was in circulation, use of ART and/or PPA together with PFGE would be useful for precise discrimination of Shigella strains.

摘要

志贺氏菌是腹泻尤其是儿童腹泻最重要的病原体之一。由于人类是志贺氏菌的主要宿主,且人与人之间可传播,志贺氏菌很容易在公共场所传播并引发疫情。在本研究中,对2001年、2008年和2009年在土耳其安卡拉分离出的60株志贺氏菌菌株进行了抗菌药敏谱、质粒图谱分析(PPA)和脉冲场凝胶电泳(PFGE)研究。为进行流行病学调查,将作为表型分析方法的抗生素耐药性分型(ART)所得结果与作为基因分型方法的PPA和PFGE结果进行了比较。在分离出的菌株中,49株(81.6%)为宋内志贺氏菌,10株(16.6%)为福氏志贺氏菌,1株(1.6%)为痢疾志贺氏菌。采用纸片扩散法评估抗菌药敏性,发现对甲氧苄啶/磺胺甲恶唑的耐药率最高(91.6%),其次是四环素(68.3%)和氨苄西林(26.6%)。福氏志贺氏菌分离株对氨苄西林、氯霉素和阿莫西林/克拉维酸的耐药性高于宋内志贺氏菌(p<0.001)。所有分离株对环丙沙星、庆大霉素和头孢他啶均敏感。宋内志贺氏菌表现出12种抗生素耐药模式,福氏志贺氏菌表现出4种。所有分离株均携带大小不同、数量在1至7个之间的质粒。宋内志贺氏菌分离株表现出27种质粒图谱,福氏志贺氏菌分离株表现出8种。通过PFGE,宋内志贺氏菌分离株聚为4种模式,福氏志贺氏菌聚为5种模式。该方法显示在安卡拉分离出的宋内志贺氏菌菌株间存在明显的克隆相似性,鉴别力(DP)计算为0.26。PPA和ART在宋内志贺氏菌菌株间显示出更高的DP(分别为0.97和0.75)。在本研究中,不稳定遗传移动元件的获得或丢失被认为是PPA和ART方法具有更高鉴别力的原因。发现这些分型方法适用于短时间内收集的菌株的流行病学调查。发现PFGE便于评估菌株的克隆相关性,然而,在同一克隆流行的此类地理区域,将ART和/或PPA与PFGE一起使用将有助于精确鉴别志贺氏菌菌株。

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