title: Hemoculture and Direct Sputum Detection of mecA-Mediated Methicillin-Resistant Staphylococcus aureus by Loop-Mediated Isothermal Amplification in Combination With a Lateral-Flow Dipstick.
abstract: This study reports loop-mediated isothermal amplification (LAMP) for rapid detection of methicillin-resistant Staphylococcus aureus from direct clinical specimens. Four primers including outer and inner primers were specifically designed on the two target sequences-femB to identify S. aureus and mecA to identify antibiotic-resistant gene. Reference strains including various species of gram-positive/gram-negative isolates were used to evaluate and optimize LAMP assays. The optimum LAMP condition was found at 63°C within 70 min assay time (include hybridization with FITC probe for 5 min and further 5 min for reading the results on the lateral flow dipstick). The detection limits of LAMP for mecA was 10 pg of total DNA or 100 CFU/ml. The LAMP assays were applied to a total of 155 samples of direct DNA extraction from sputum and hemoculture bottles. The sensitivity of LAMP for mecA detection in sputum and hemoculture bottles was 93.3% (28/30) and 100% (52/52), respectively. In conclusion, LAMP assay is an alternative technique for rapid detection of MRSA infection with a technical simplicity and cost-effective method in a routine diagnostic laboratory.
authors.affiliation: Materials Science and Engineering Programme, Faculty of Science, Mahidol University, Bangkok, Thailand.
authors.forename: Kawin
authors.initials: K
authors.lastname: Nawattanapaiboon
authors.affiliation: Materials Science and Engineering Programme, Faculty of Science, Mahidol University, Bangkok, Thailand.
authors.forename: Photchanathorn
authors.initials: P
authors.lastname: Prombun
authors.affiliation: Department of Pathology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Pitak
authors.initials: P
authors.lastname: Santanirand
authors.affiliation: Department of Pathology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Apirom
authors.initials: A
authors.lastname: Vongsakulyanon
authors.affiliation: Materials Science and Engineering Programme, Faculty of Science, Mahidol University, Bangkok, Thailand.|Department of Physics, Faculty of Science, Mahidol University, Bangkok, Thailand.
authors.forename: Toemsak
authors.initials: T
authors.lastname: Srikhirin
authors.affiliation: School of Electronic Engineering, Institute of Engineering, , Suranaree University of Technology, Nakhon Ratchasima, Thailand.
authors.forename: Boonsong
authors.initials: B
authors.lastname: Sutapun
authors.affiliation: Bioengineering and Sensing Technology Laboratory, BIOTEC, National Science and Technology Development Agency (NSTDA), Pathum Thani, Thailand. wansika@biotec.or.th.
authors.forename: Wansika
authors.initials: W
authors.lastname: Kiatpathomchai
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D001483:Base Sequence; D001710:Biotin / Q000378:metabolism; D017931:DNA Primers / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D005798:Genes, Bacterial; D006801:Humans; D057230:Limit of Detection; D055624:Methicillin-Resistant Staphylococcus aureus / Q000235:genetics* / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012212:Rheology; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology*
chemical_list: D017931:DNA Primers; D001710:Biotin
keywords: MRSA; clinical specimens; lateral flow dipstick; loop-mediated isothermal amplification; methicillin-resistant Staphylococcus aureus