4.8 Article

SPR-DNA array for detection of methicillin-resistant Staphylococcus aureus (MRSA) in combination with loop-mediated isothermal amplification

Journal

BIOSENSORS & BIOELECTRONICS
Volume 74, Issue -, Pages 335-340

Publisher

ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2015.06.038

Keywords

Surface plasmon resonance; Loop-mediated isothermal amplification; DNA array; DNA sensor; Methicillin-resistant Staphylococcus aureus; Clinical specimen

Funding

  1. National for Electronics and Computer Technology Center (NECTEC), Laboratory of Clinical Microbiology, Department of Pathology, Ramathibodi Hospital
  2. National Center for Genetic Engineering and Biotechnology (BIOTEC)
  3. Center of Excellence (COE) of National Nanotechnology Center (NANOTEC)

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In this study, we evaluated surface plasmon resonance imaging (SPR imaging) as a DNA biosensor for the detection of methicillin-resistant Staphylococcus aureus (MRSA) which is one of the most common causes of nosocomial infections. The DNA sample were collected from clinical specimens, including sputum and blood hemoculture were undergone LAMP amplification for 0.18 kbp and 0.23 kbp DNA fragments of femB and mecA genes, respectively. The self-assembled monolayer surface (SAMs) was used for immobilized streptavidin-biotinylated probes on the sensor surface for the detection of LAMP amplicons from MRSA. Both LAMP amplicons were simultaneously hybridized with ssDNA probes immobilized onto a bio-functionalized surface to detect specific targets in the multiplex DNA array platform. In addition, the sensor surface could be regenerated allowing at least five cycles of use with a shortened assay time. The detection limit of LAMP-SPR sensing was 10 copies/RI and LAMP-SPR sensing system showed a good selectivity toward the MRSA. (C) 2015 Elsevier B.V. All rights reserved.

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