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Hemoculture and Direct Sputum Detection of mecA-Mediated Methicillin-Resistant Staphylococcus aureus by Loop-Mediated Isothermal Amplification in Combination With a Lateral-Flow Dipstick
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Document 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
Author
Nawattanapaiboon K., Prombun P., Santanirand P., Vongsakulyanon A., Srikhirin T., Sutapun B., Kiatpathomchai W.
Name from Authors Collection
Affiliations
Materials Science and Engineering Programme, Faculty of Science, Mahidol UniversityBangkok, Thailand; Department of Pathology, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityBangkok, Thailand; Department of Physics, Faculty of Science, Mahidol UniversityBangkok, Thailand; School of Electronic Engineering, Institute of Engineering, Suranaree University of Technology, Nakhon Ratchasima, Thailand; Bioengineering and Sensing Technology Laboratory, BIOTEC, National Science and Technology Development Agency (NSTDA), Pathum Thani, Thailand
Type
Article
Source Title
Journal of Clinical Laboratory Analysis
ISSN
08878013
Year
2016
Volume
30
Issue
5
Page
760-767
Open Access
Green
Publisher
John Wiley and Sons Inc.
DOI
10.1002/jcla.21935
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. © 2016 Wiley Periodicals, Inc.
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
License
CC BY, a CC BY-NC or a CC BY-NC-ND
Rights
Wiley, China
Publication Source
Scopus