Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA) |
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| Affiliation: | 1. Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia;2. Department of Medical Microbiology, Basrah University, Basrah, Iraq;3. Laboratory of Marine Science and Aquaculture, Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia;4. La Balme Microbiology Unit, BioMérieux, 3 route de Port Michaud, 38390 La Balme-les-Grottes, France;5. Medical Genetics Laboratory, Department of Obstetrics & Gynaecology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia;1. Infectious Diseases Service, University Hospital Centre and University of Lausanne, Lausanne, Switzerland;2. Institute of Microbiology, University Hospital Centre and University of Lausanne, Lausanne, Switzerland;3. Hospital Preventive Medicine Service, University Hospital Centre and University of Lausanne, Lausanne, Switzerland;1. Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine and Zoothecny, University of São Paulo (USP), Av. Prof. Dr. Orlando Marques de Paiva, 87, CEP 05508-270, São Paulo, Brazil;2. Federal University of Piauí (UFPI), Campus Senador Helvídio Nunes de Barros, Rua Cícero Eduardo, S/N, Junco, CEP 64600-000, Picos, Piauí, Brazil;3. Institute of Biomedical Sciences, University of São Paulo (USP), Av. Prof. Lineu Prestes, 1374 – Ed. Biomédicas II, Sala 240, Butantã, CEP 05508-900, São Paulo, Brazil;1. Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran;2. Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran;3. Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran;4. Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, Iran;1. Department of Microbiology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran;2. Department of Microbiology, Faculty of Medicine, Golestan University of Medical Sciences, Gorgan, Iran;3. Department of Microbiology, Faculty of Medicine, Shahid Beheshti University of Medical Science, Tehran, Iran;4. Department of Pediatric Infectious Disease, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran;5. Department of Microbiology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran;6. Department of Pharmacology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran;1. Laboratorio de Biología Celular y Molecular Aplicada, Instituto de Ciencias Veterinarias del Litoral (ICIVET-Litoral), Universidad Nacional del Litoral (UNL)/Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Esperanza, Santa Fe, Argentina;2. Departamento de Ciencias Morfológicas, Facultad de Ciencias Veterinarias, Universidad Nacional del Litoral (UNL), Esperanza, Santa Fe, Argentina;3. Laboratorio de Biología Molecular e Inmunología Aplicadas, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral (UNL), Santa Fe, Argentina;4. Estación Experimental Agropecuaria Rafaela, Instituto Nacional de Tecnología Agropecuaria (INTA), Rafaela, Santa Fe, Argentina |
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| Abstract: | Staphylococcus aureus biofilm associated infections remains a major clinical concern in patients with indwelling devices. Quantitative real-time PCR (qPCR) can be used to investigate the pathogenic role of such biofilms. We describe qPCRs for 12 adhesion and biofilm-related genes of four S. aureus isolates which were applied during in vitro biofilm development. An endogenous control (16S rRNA) was used for signal normalization. We compared the qPCR results with structural analysis using scanning electron microscopy (SEM). The SEM studies showed different cellular products surrounding the aggregated cells at different times of biofilm formation. Using qPCR, we found that expression levels of the gene encoding fibronectin binding protein A and B and clumping factor B (fnbA/B and clfB), which involves in primary adherence of S. aureus, were significantly increased at 24 h and decreased slightly and variably at 48 h when all 4 isolates were considered. The elastin binding protein (ebps) RNA expression level was significantly enhanced more than 6-fold at 24 and 48 h compared to 12 h. Similar results were obtained for the intercellular adhesion biofilm required genes type C (icaC). In addition, qPCR revealed a fluctuation in expression levels at different time points of biofilm growth of other genes, indicating that different parameter modes of growth processes are operating at different times. |
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| Keywords: | Biofilm SEM qPCR Adhesion Biofilm related gene expression |
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