Rapid Isolation of Staphylococcus aureus Pathogens from Infected Clinical Samples Using Magnetic Beads Coated with Fc-Mannose Binding Lectin
| Autor: | Michael Super, M. Penary, Benjamin T. Seiler, Donald E. Ingber, Mark Cartwright, Melissa Rodas, Martin Rottman, Eric Oswald, Nazita Gamini, G. Giordano, A. Bicart-See |
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| Rok vydání: | 2016 |
| Předmět: |
Male
0301 basic medicine Interstitial Fluid Physiology Staphylococcus lcsh:Medicine Pathology and Laboratory Medicine medicine.disease_cause Biochemistry Immune Physiology Synovial Fluid Medicine and Health Sciences lcsh:Science Mannan-binding lectin Immune System Proteins Multidisciplinary biology Organic Compounds Proteases Microspheres Bacterial Pathogens Enzymes Body Fluids Chemistry Medical Microbiology Staphylococcus aureus Physical Sciences Female Sample collection Pathogens Anatomy Research Article Recombinant Fusion Proteins Antibiotic sensitivity Immunology Carbohydrates Mannose-Binding Lectin Microbiology Antibodies 03 medical and health sciences medicine Humans Synovial fluid Microbial Pathogens Opsonin Bacteria 030102 biochemistry & molecular biology lcsh:R Organic Chemistry Organisms Chemical Compounds Biology and Life Sciences Proteins biology.organism_classification Immunoglobulin Fc Fragments Magnetic Fields 030104 developmental biology Enzymology lcsh:Q |
| Zdroj: | PLoS ONE PLoS ONE, Vol 11, Iss 6, p e0156287 (2016) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0156287 |
| Popis: | Here we describe how Staphylococcus aureus bacteria can be rapidly isolated from clinical samples of articular fluid and synovial tissue using magnetic beads coated with the engineered chimeric human opsonin protein, Fc-mannose-binding lectin (FcMBL). The FcMBL-beads were used to capture and magnetically remove bacteria from purified cultures of 12 S. aureus strains, and from 8 articular fluid samples and 4 synovial tissue samples collected from patients with osteoarthritis or periprosthetic infections previously documented by positive S. aureus cultures. While the capture efficiency was high (85%) with purified S. aureus strains grown in vitro, direct FcMBL-bead capture from the clinical samples was initially disappointing (< 5% efficiency). Further analysis revealed that inhibition of FcMBL binding was due to coating of the bacteria by immunoglobulins and immune cells that masked FcMBL binding sites, and to the high viscosity of these complex biological samples. Importantly, capture of pathogens using the FcMBL-beads was increased to 76% efficiency by pretreating clinical specimens with hypotonic washes, hyaluronidase and a protease cocktail. Using this approach, S. aureus bacteria could be isolated from infected osteoarthritic tissues within 2 hours after sample collection. This FcMBL-enabled magnetic method for rapid capture and concentration of pathogens from clinical samples could be integrated upstream of current processes used in clinical microbiology laboratories to identify pathogens and perform antibiotic sensitivity testing when bacterial culture is not possible or before colonies can be detected. |
| Databáze: | OpenAIRE |
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