Crawford Evan C, Singh Ameet, Gibson Thomas W G, Scott Weese J
Department of Clinical Studies.
Centre for Public Health and Zoonoses, Department of Pathology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada.
Vet Surg. 2016 May;45(4):499-506. doi: 10.1111/vsu.12471. Epub 2016 Apr 15.
To evaluate the expression of biofilm-associated genes in Staphylococcus pseudintermedius on multiple clinically relevant surfaces.
In vitro experimental study.
Two strains of methicillin-resistant S. pseudintermedius isolated from clinical infections representing the most common international isolates.
A quantitative polymerase chain reaction (qPCR) assay for expression of genes related to biofilm initial adhesion, formation/maturation, antimicrobial resistance, and intracellular communication was developed and validated. S. pseudintermedius biofilms were grown on 8 clinically relevant surfaces (polymethylmethacrylate, stainless steel, titanium, latex, silicone, polydioxanone, polystyrene, and glass) and samples of logarithmic and stationary growth phases were collected. Gene expression in samples was measured by qPCR.
Significant differences in gene expression were identified between surfaces and between bacterial strains for most gene/strain/surface combinations studied. Expression of genes responsible for production of extracellular matrix were increased in biofilms. Expression of genes responsible for initial adhesion and intracellular communication was markedly variable. Antimicrobial resistance gene expression was increased on multiple surfaces, including stainless steel and titanium.
A method for evaluation of expression of multiple biofilm-associated genes in S. pseudintermedius was successfully developed and applied to the study of biofilms on multiple surfaces. Variations in expression of these genes have a bearing on understanding the development and treatment of implant-associated biofilm infections and will inform future clinical research.
评估多株中间型葡萄球菌在多种临床相关表面上生物膜相关基因的表达情况。
体外实验研究。
从临床感染中分离出的两株耐甲氧西林中间型葡萄球菌,代表了国际上最常见的分离菌株。
开发并验证了一种用于检测与生物膜初始黏附、形成/成熟、抗菌耐药性及细胞间通讯相关基因表达的定量聚合酶链反应(qPCR)检测方法。中间型葡萄球菌生物膜在8种临床相关表面(聚甲基丙烯酸甲酯、不锈钢、钛、乳胶、硅胶、聚二氧六环酮、聚苯乙烯和玻璃)上生长,并收集对数生长期和稳定生长期的样本。通过qPCR测定样本中的基因表达。
在所研究的大多数基因/菌株/表面组合中,表面之间以及菌株之间的基因表达存在显著差异。生物膜中负责细胞外基质产生的基因表达增加。负责初始黏附和细胞间通讯的基因表达明显可变。包括不锈钢和钛在内的多种表面上抗菌耐药基因的表达增加。
成功开发了一种评估中间型葡萄球菌中多种生物膜相关基因表达的方法,并将其应用于多种表面生物膜的研究。这些基因表达的变化与理解植入物相关生物膜感染的发生发展及治疗有关,并将为未来的临床研究提供信息。