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

Kawin Nawattanapaiboon; Wansika Kiatpathomchai; Pitak Santanirand; Apirom Vongsakulyanon; Ratthasart Amarit; Armote Somboonkaew; Boonsong Sutapun; Toemsak Srikhirin

doi:10.1016/j.bios.2015.06.038

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

Biosens Bioelectron. 2015 Dec 15:74:335-40. doi: 10.1016/j.bios.2015.06.038. Epub 2015 Jun 25.

Authors

Kawin Nawattanapaiboon¹, Wansika Kiatpathomchai², Pitak Santanirand³, Apirom Vongsakulyanon³, Ratthasart Amarit⁴, Armote Somboonkaew⁴, Boonsong Sutapun⁵, Toemsak Srikhirin⁶

Affiliations

¹ Materials Science and Engineering Programme, Faculty of Science, Mahidol University, Rama VI Rd., Phayathai, Bangkok 10400, Thailand.
² Bioengineering and Sensing Technology Laboratory, BIOTEC, National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.
³ Department of Pathology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Rama VI Rd., Phayathai, Bangkok 10400, Thailand.
⁴ Photonic Technology Laboratory, NECTEC, National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, Thailand.
⁵ School of Electronic Engineering, Institute of Engineering, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand.
⁶ 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.

PMID: 26159153
DOI: 10.1016/j.bios.2015.06.038

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.

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

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Load / instrumentation
Base Sequence
Biosensing Techniques / instrumentation
DNA, Bacterial / genetics*
DNA, Bacterial / isolation & purification
Equipment Design
Equipment Failure Analysis
Humans
Methicillin-Resistant Staphylococcus aureus / genetics*
Methicillin-Resistant Staphylococcus aureus / isolation & purification*
Molecular Sequence Data
Nucleic Acid Amplification Techniques / instrumentation*
Oligonucleotide Array Sequence Analysis / instrumentation*
Reproducibility of Results
Sensitivity and Specificity
Surface Plasmon Resonance / instrumentation*

Substances

DNA, Bacterial