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

Nawattanapaiboon, Kawin, Kiatpathomchai, Wansika, Santanirand, Pitak, Vongsakulyanon, Apirom, Amarit, Ratthasart, Somboonkaew, Armote, Sutapun, Boonsong, Srikhirin, Toemsak
Biosensors & bioelectronics 2015 v.74 pp. 335-340
DNA fragmentation, biosensors, blood, cross infection, detection limit, genes, image analysis, loop-mediated isothermal amplification, methicillin, methicillin-resistant Staphylococcus aureus, single-stranded DNA, surface plasmon resonance
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 0.18kbp and 0.23kbp 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/µl and LAMP–SPR sensing system showed a good selectivity toward the MRSA.