title: SPR-DNA array for detection of methicillin-resistant Staphylococcus aureus (MRSA) in combination with loop-mediated isothermal amplification.
abstract: 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/µl and LAMP-SPR sensing system showed a good selectivity toward the MRSA.
authors.affiliation: Materials Science and Engineering Programme, Faculty of Science, Mahidol University, Rama VI Rd., Phayathai, Bangkok 10400, Thailand.
authors.forename: Kawin
authors.initials: K
authors.lastname: Nawattanapaiboon
authors.affiliation: Bioengineering and Sensing Technology Laboratory, BIOTEC, National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.
authors.forename: Wansika
authors.initials: W
authors.lastname: Kiatpathomchai
authors.affiliation: Department of Pathology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Rama VI Rd., Phayathai, Bangkok 10400, Thailand.
authors.forename: Pitak
authors.initials: P
authors.lastname: Santanirand
authors.affiliation: Department of Pathology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Rama VI Rd., Phayathai, Bangkok 10400, Thailand.
authors.forename: Apirom
authors.initials: A
authors.lastname: Vongsakulyanon
authors.affiliation: Photonic Technology Laboratory, NECTEC, National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.
authors.forename: Ratthasart
authors.initials: R
authors.lastname: Amarit
authors.affiliation: Photonic Technology Laboratory, NECTEC, National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.
authors.forename: Armote
authors.initials: A
authors.lastname: Somboonkaew
authors.affiliation: School of Electronic Engineering, Institute of Engineering, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand.
authors.forename: Boonsong
authors.initials: B
authors.lastname: Sutapun
authors.affiliation: Materials Science and Engineering Programme, Faculty of Science, Mahidol University, Rama VI Rd., Phayathai, Bangkok 10400, Thailand; Physics Department, Faculty of Science, Mahidol University, Rama VI Rd., Phayathai, Bangkok 10400, Thailand. Electronic address: sctsk@mahidol.ac.th.
authors.forename: Toemsak
authors.initials: T
authors.lastname: Srikhirin
mesh_terms: D058491:Bacterial Load / Q000295:instrumentation; D001483:Base Sequence; D015374:Biosensing Techniques / Q000295:instrumentation; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000235:genetics* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000295:instrumentation*
chemical_list: D004269:DNA, Bacterial
keywords: Clinical specimen; DNA array; DNA sensor; Loop-mediated isothermal amplification; Methicillin-resistant Staphylococcus aureus; Surface plasmon resonance