Design and Development of Geno-Chromatography Based Biosensor Assay for the Rapid Identification of vanA Gene (original) (raw)

Abstract

Background: Glycopeptide-resistant enterococci (GRE) have emerged as a significant cause of nosocomial infections worldwide due to widespread use of glycopeptide antimicrobial drugs such as vancomycin, teicoplanin in hospitals and avorporcin as a growth promoter in poultry industry. A fast and reliable on-site monitoring scheme is required for quick preventive action and early diagnosis. Hence we have developed a rapid geno-chromatography biosensor assay for the identification of the high-level vancomycin- and teicoplanin-resistance gene, vanA. Methods: vanA PCR was performed with biotin- and fluorescein-labeled primers. A redox current signal was generated by the action of peroxidase-conjugated anti-fluorescein antibody with hydrogen peroxide and was detected using a combination of glass membrane filter chromatography and electrochemical DNA biosensor on a portable amperometer. An internal control was incorporated for assay reliability. The specificity and sensitivity was evaluated...

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