Kawin NawattanapaiboonPhotchanathorn PrombunPitak SantanirandApirom VongsakulyanonToemsak SrikhirinBoonsong SutapunWansika KiatpathomchaiMahidol UniversitySuranaree University of TechnologyThailand National Center for Genetic Engineering and Biotechnology2018-12-112019-03-142018-12-112019-03-142016-09-01Journal of Clinical Laboratory Analysis. Vol.30, No.5 (2016), 760-76710982825088780132-s2.0-84989245039https://repository.li.mahidol.ac.th/handle/20.500.14594/42897© 2016 Wiley Periodicals, Inc. 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.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyHealth ProfessionsMedicineArticleSCOPUS10.1002/jcla.21935