Evaluation of the occurrence of genetic determinants of multi-drug resistance among Acinetobacter baumannii strains

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Abstract

Introduction: The aim of the study was the analysis of occurrence of genetic determinants of multi-drug resistance and the assessment of genetic relationship among Acinetobacter baumannii strains. Methods: Multiplex-PCR method was performed in order to: (1) confirm the phenotypic identification and (2) detect the presence of CHDL oxacillinases in the group of thirty A.baumannii strains. Further PCR studies included the analysis of the occurrence of genetic determinants associated with efflux pump, insertion sequence and biofilm formation. The relationship between bacterial strains was assayed using 6 primers in RAPD-PCR method. Results: Detection of the blaOXA-51-like gene confirmed that the strains belong to the A. baumannii species. In the multiplex-PCR, the presence of the blaOXA-23-like and blaOXA-40-like genes was detected in 3 (10%) and 27 (90%) isolates, respectively. Moreover, some strains showed the coexistence of the blaOXA-51-like and blaOXA-23-like genes (10%, n=3) or blaOXA-51-like and blaOXA-40-like (90%, n=27). In the group of analysed strains the presence of the efflux pump gene (adeA) and the insertion sequence ISAba1 were demonstrated in all tested isolates. Biofilm-related genes (abaI, csuE) were found in 100% and 97% (n=29) tested strains adequately. The RAPD-PCR studies revealed the presence of 10 unrelated genotypes. Conclusions: The obtained results suggest that the phenomenon of multi-drug resistance in the studied A. baumannii strains could be attributed to the occurrence of CHDL oxacillinases, AdeABC efflux pump, insertion sequence ISAba1 and the biofilm formation.
鲍曼不动杆菌多药耐药遗传因素的评价
前言:本研究的目的是分析鲍曼不动杆菌多药耐药遗传决定因素的发生情况,并评估鲍曼不动杆菌各菌株间的遗传关系。方法:采用多重聚合酶链反应(multiplexx - pcr)方法:(1)证实表型鉴定;(2)检测30株鲍曼不动杆菌中是否存在CHDL oxacillinases。进一步的PCR研究包括分析与外排泵、插入序列和生物膜形成相关的遗传决定因素的发生。采用RAPD-PCR方法,用6条引物分析菌株间的关系。结果:blaoxa -51样基因检测证实该菌株属于鲍曼不动杆菌属。在多重pcr中,分别在3株(10%)和27株(90%)分离株中检测到blaoxa -23和blaoxa -40样基因。部分菌株同时存在blaOXA-51-like和blaOXA-23-like基因(10%,n=3)或blaOXA-51-like和blaOXA-40-like基因(90%,n=27)。在所分析的菌株中,所有分离株均存在外排泵基因(adeA)和插入序列ISAba1。生物膜相关基因(abaI, csuE)分别在100%和97% (n=29)菌株中被充分检出。RAPD-PCR研究显示存在10个不相关的基因型。结论:本研究结果提示鲍曼不动杆菌多重耐药现象可能与CHDL oxacillinases、AdeABC外排泵、插入序列ISAba1以及生物膜的形成有关。
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