Gomila Margarita, Prince-Manzano Claudia, Svensson-Stadler Liselott, Busquets Antonio, Erhard Marcel, Martínez Deny L, Lalucat Jorge, Moore Edward R B
Microbiology, Department of Biology, University of the Balearic Islands, Palma de Mallorca, Islas Baleares, Spain.
Department of Infectious Diseases, Culture Collection University of Gothenburg (CCUG), Sahlgrenska Academy of the University of Gothenburg, Gothenburg, Sweden.
PLoS One. 2014 Dec 4;9(12):e114356. doi: 10.1371/journal.pone.0114356. eCollection 2014.
The Achromobacter is a genus in the family Alcaligenaceae, comprising fifteen species isolated from different sources, including clinical samples. The ability to detect and correctly identify Achromobacter species, particularly A. xylosoxidans, and differentiate them from other phenotypically similar and genotypically related Gram-negative, aerobic, non-fermenting species is important for patients with cystic fibrosis (CF), as well as for nosocomial and other opportunistic infections. Traditional phenotypic profile-based analyses have been demonstrated to be inadequate for reliable identifications of isolates of Achromobacter species and genotypic-based assays, relying upon comparative 16S rRNA gene sequence analyses are not able to insure definitive identifications of Achromobacter species, due to the inherently conserved nature of the gene. The uses of alternative methodologies to enable high-resolution differentiation between the species in the genus are needed. A comparative multi-locus sequence analysis (MLSA) of four selected 'house-keeping' genes (atpD, gyrB, recA, and rpoB) assessed the individual gene sequences for their potential in developing a reliable, rapid and cost-effective diagnostic protocol for Achromobacter species identifications. The analysis of the type strains of the species of the genus and 46 strains of Achromobacter species showed congruence between the cluster analyses derived from the individual genes. The MLSA gene sequences exhibited different levels of resolution in delineating the validly published Achromobacter species and elucidated strains that represent new genotypes and probable new species of the genus. Our results also suggested that the recently described A. spritinus is a later heterotypic synonym of A. marplatensis. Strains were analyzed, using whole-cell Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight mass spectrometry (MALDI-TOF MS), as an alternative phenotypic profile-based method with the potential to support the identifications determined by the genotypic DNA sequence-based MLSA. The MALDI-TOF MS data showed good accordance in strain groupings and identifications by the MLSA data.
无色杆菌属是产碱菌科中的一个属，包含从不同来源（包括临床样本）分离出的15个种。对于囊性纤维化（CF）患者以及医院内感染和其他机会性感染而言，检测并正确鉴定无色杆菌属菌种，尤其是木糖氧化无色杆菌，并将其与其他表型相似且基因相关的革兰氏阴性、需氧、非发酵菌种区分开来非常重要。已证明基于传统表型特征的分析不足以可靠鉴定无色杆菌属菌种的分离株，而基于基因分型的检测方法（依靠比较16S rRNA基因序列分析）由于该基因固有的保守性质，也无法确保对无色杆菌属菌种进行明确鉴定。因此需要使用替代方法来实现该属内各菌种之间的高分辨率区分。对四个选定的“管家”基因（atpD、gyrB、recA和rpoB）进行的比较多位点序列分析（MLSA）评估了各个基因序列在开发一种可靠、快速且经济高效的无色杆菌属菌种鉴定诊断方案方面的潜力。对该属各菌种的模式菌株和46株无色杆菌属菌株进行分析，结果表明来自各个基因的聚类分析之间具有一致性。MLSA基因序列在划分已有效发表的无色杆菌属菌种以及阐明代表该属新基因型和可能新物种的菌株方面表现出不同程度的分辨率。我们的结果还表明，最近描述的灵活动无色杆菌是马尔普拉无色杆菌的晚出异名。使用全细胞基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）对菌株进行分析，作为一种基于表型特征的替代方法，有可能支持基于基因型DNA序列的MLSA所确定的鉴定结果。MALDI-TOF MS数据在菌株分组和鉴定方面与MLSA数据显示出良好的一致性。
