Design and Development of a Novel Electrochemical DNA Biosensor for Rapid Molecular Identification of Enterococcus faecium
Makaleler > Tam Makale · 2008
Yazarlar: Y.Y. Chan, K. Balqis, A.O. Dilsat, S.Y. Lee, P. Lalitha, M. Ozsoz, M. Ravichandran
International Journal of Infectious Diseases, Uluslararası, Wos, 2008, cilt 12, ss e59-e59
DOI: 10.1016/j.ijid.2008.05.037
Özet
Enterococci have emerged as prominent nosocomial pathogens that cause a variety of clinical infections in many parts of the world over the last decade. The most common enterococci strains isolated from clinical samples are E. faecium and E. faecalis. Enterococci are known to have acquired resistance to vancomycin (glycopeptide) antimicrobials, resulting in the rapid increase of vancomycin resistant enterococci (VRE) strains in human. The conventional culture methods are time-consuming and laborious. Alternative molecular techniques polymerase chain reaction (PCR) and agarose gel electrophoresis utilize harmful elements such as carcinogenic ultraviolet light and ethidium bromide. In addition, optical-based techniques such as real-time PCR are expensive and require specialized equipments. Recently, interest has been increasing in the development of simple, inexpensive and disposable DNA biosensors for field and clinical assays. In the present study, an electrochemical DNA biosensor was designed and developed for detection of E. faecium. Design, fabrication and electrochemical characterization of screen-printed carbon electrodes (SPCEs) were carried-out. Optimization of the PCR and biosensor protocols such as PCR hapten labeling, washing step and peroxidase oxidation signal were performed. Under the optimized conditions, the oxidation signal threshold value was determined at 2.00 ± 0.02 μA. The analytical specificity of the biosensor assay was evaluated with reference E. faecium and non-E. faecium strains and was found to be 100%, while analytical sensitivity of the assay was 10 CFU/ml. The biosensor assay gave quantitative results rather then qualitative results when compared with agarose gel and DNA-chromatography based tests. In this study, the biosensor was optimized using E. faecium as a model organism and proved to be sensitive and specific. Hence in future, it will be possible to use this biosensor for antimicrobial resistant determinants, other microorganisms or mutant gene detection in hospital and environmental settings. 13th International Congress on Infectious Diseases Abstracts (Oral Presentations)International Journal of Infectious DiseasesVol. 12Preview Full-Text PDF Open Archive