Publication: Hemoculture and Direct Sputum Detection of mecA-Mediated Methicillin-Resistant Staphylococcus aureus by Loop-Mediated Isothermal Amplification in Combination With a Lateral-Flow Dipstick
Issued Date
2016-09-01
Resource Type
ISSN
10982825
08878013
08878013
Other identifier(s)
2-s2.0-84989245039
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Clinical Laboratory Analysis. Vol.30, No.5 (2016), 760-767
Suggested Citation
Kawin Nawattanapaiboon, Photchanathorn Prombun, Pitak Santanirand, Apirom Vongsakulyanon, Toemsak Srikhirin, Boonsong Sutapun, Wansika Kiatpathomchai Hemoculture and Direct Sputum Detection of mecA-Mediated Methicillin-Resistant Staphylococcus aureus by Loop-Mediated Isothermal Amplification in Combination With a Lateral-Flow Dipstick. Journal of Clinical Laboratory Analysis. Vol.30, No.5 (2016), 760-767. doi:10.1002/jcla.21935 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/42897
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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
© 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.