Sousa Telma de, Viala Didier, Théron Laetitia, Chambon Christophe, Hébraud Michel, Poeta Patricia, Igrejas Gilberto
Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal.
Microbiology and Antibiotic Resistance Team (MicroART), Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal.
Biology (Basel). 2020 Mar 19;9(3):56. doi: 10.3390/biology9030056.
The commensal bacteria causes several intestinal and extra-intestinal diseases, since it has virulence factors that interfere in important cellular processes. These bacteria also have a great capacity to spread the resistance genes, sometimes to phylogenetically distant bacteria, which poses an additional threat to public health worldwide. Here, we aimed to use the analytical potential of MALDI-TOF mass spectrometry (MS) to characterize isolates and identify proteins associated closely with antibiotic resistance. Thirty strains of extended-spectrum beta-lactamase producing were sampled from various animals. The phenotypes of antibiotic resistance were determined according to Clinical and Laboratory Standards Institute (CLSI) methods, and they showed that all bacterial isolates were multi-resistant to trimethoprim-sulfamethoxazole, tetracycline, and ampicillin. To identify peptides characteristic of resistance to particular antibiotics, each strain was grown in the presence or absence of the different antibiotics, and then proteins were extracted from the cells. The protein fingerprints of the samples were determined by MALDI-TOF MS in linear mode over a mass range of 2 to 20 kDa. The spectra obtained were compared by using the ClinProTools bioinformatics software, using three machine learning classification algorithms. A putative species biomarker was also detected at a peak / of 4528.00.
共生细菌会引发多种肠道和肠道外疾病,因为它具有干扰重要细胞过程的毒力因子。这些细菌还具有强大的传播耐药基因的能力,有时会传播到系统发育关系较远的细菌,这对全球公共卫生构成了额外威胁。在此,我们旨在利用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)的分析潜力来表征分离株,并鉴定与抗生素耐药性密切相关的蛋白质。从各种动物中采集了30株产超广谱β-内酰胺酶的菌株。根据临床和实验室标准协会(CLSI)的方法确定抗生素耐药表型,结果显示所有细菌分离株对甲氧苄啶-磺胺甲恶唑、四环素和氨苄西林均具有多重耐药性。为了鉴定对特定抗生素耐药的特征性肽段,将每个菌株分别在存在或不存在不同抗生素的情况下培养,然后从细胞中提取蛋白质。通过MALDI-TOF MS以线性模式在2至20 kDa的质量范围内测定样品的蛋白质指纹图谱。使用三种机器学习分类算法,通过ClinProTools生物信息学软件对获得的光谱进行比较。在4528.00的峰/处还检测到一个假定的物种生物标志物。