Drożdż Kamil, Ochońska Dorota, Ścibik Łukasz, Gołda-Cępa Monika, Biegun Katarzyna, Brzychczy-Włoch Monika
Department of Molecular Medical Microbiology, Chair of Microbiology, Faculty of Medicine, Jagiellonian University Medical College, 31-121 Krakow, Poland.
Department of Otolaryngology, Head and Neck Surgery, 5th Military Hospital with Polyclinic, 30-090 Krakow, Poland.
Microorganisms. 2022 Jun 14;10(6):1210. doi: 10.3390/microorganisms10061210.
: Bacterial biofilm on the surface of tracheostomy tubes (TTs) is a potential reservoir of potentially pathogenic bacteria, including . For this reason, our study aimed to investigate biofilm production in vitro and the presence of AD and MSCRAMM genes in clinical strains derived from TTs, with respect to antibiotic resistance and genetic variability. The clonality of the strains was analyzed by the PFGE method. The assessment of drug resistance was based on the EUCAST recommendations. The isolates were evaluated for biofilm production by the microtiter plate method and the slime-forming ability was tested on Congo red agar (CRA). The presence of AD genes was investigated by PCR and MSCRAMM genes were detected by multiplex PCR. A total of 60 patients were enrolled in the study. One TT was obtained from each patient (n = 60). Twenty-one TTs (35%) were colonized with S. aureus. A total of 24 strains were isolated as 3 patients showed colonization with 2 SA clones (as confirmed by PFGE). PFGE showed twenty-two unique molecular profiles. Two isolates (8%) turned out to be MRSA, but 50% were resistant to chloramphenicol, 25% to erythromycin and 8% to clindamycin (two cMLS and four iMLS phenotypes were detected). The microtiter plate method with crystal violet confirmed that 96% of the strains were biofilm formers. Representative strains were visualized by SEM. All isolates had AB, A, S and AD. Different MSCRAMM gene combinations were observed. the present study showed that the isolated from the TTs has a high diversity of genotypes, a high level of antibiotic resistance and ability to produce biofilm.
气管造口管(TTs)表面的细菌生物膜是潜在病原菌的一个潜在储存库,包括……。因此,我们的研究旨在调查TTs临床分离株的体外生物膜形成情况以及粘附素(AD)和微生物表面成分识别黏附分子(MSCRAMM)基因的存在情况,同时研究其抗生素耐药性和遗传变异性。通过脉冲场凝胶电泳(PFGE)方法分析分离株的克隆性。耐药性评估基于欧洲抗菌药物敏感性试验委员会(EUCAST)的建议。采用微量滴定板法评估分离株的生物膜形成情况,并在刚果红琼脂(CRA)上测试其产黏液能力。通过聚合酶链反应(PCR)研究AD基因的存在情况,通过多重PCR检测MSCRAMM基因。共有60名患者纳入本研究。从每位患者获取一根TT(n = 60)。21根TT(35%)被金黄色葡萄球菌定植。共分离出24株菌株,因为有3名患者显示被2个金黄色葡萄球菌克隆定植(经PFGE确认)。PFGE显示出22种独特的分子图谱。2株分离株(8%)为耐甲氧西林金黄色葡萄球菌(MRSA),但50%对氯霉素耐药,25%对红霉素耐药,8%对克林霉素耐药(检测到2种cMLS和4种iMLS表型)。用结晶紫的微量滴定板法证实96%的菌株为生物膜形成菌。通过扫描电子显微镜(SEM)观察代表性菌株。所有分离株都有AB、A、S和AD。观察到不同的MSCRAMM基因组合。本研究表明,从TTs分离出的菌株具有高度的基因型多样性、高水平的抗生素耐药性和生物膜形成能力。