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Application of Loop-Mediated Isothermal Amplification (LAMP) Assay to Rapid Detection of Methicillin-Resistant Staphylococcus aureus from Blood Cultures
Biomed Sci Letters 2019;25:75-82
Published online March 31, 2019;  https://doi.org/10.15616/BSL.2019.25.1.75
© 2019 The Korean Society For Biomedical Laboratory Sciences.

Yun-Hee Baek1,짠,*, Mi-Young Jo2,3,짠,*, Min-Suk Song1,*,Seung-Bok Hong4,* and Kyeong-Seob Shin2,,*

Department of Microbiology1, Laboratory Medicine2, Chungbuk National University College of Medicine, Cheongju 28644, Korea 3Department of Nursing Science, Kyungbuk College of Natural Sciences, Yeongju 36133, Korea 4Department of Clinical Laboratory Science, Chungbuk Health & Science Unviersity, Cheongju 28150, Korea
Correspondence to: Kyeong-Seob Shin. Department of Laboratory Medicine, Chungbuk National University College of Medicine, Cheong-ju 28644, Korea.
Tel: +82-43-269-6240, Fax: +82-43-271-5243, e-mail: ksshin@chungbuk.ac.kr
Received February 8, 2019; Revised March 7, 2019; Accepted March 19, 2019.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
 Abstract
We developed the multiplex LAMP assay using 16S rRNA, femA and mecA genes for direct detection of the methicillin resistance in Staphylococci from positive blood culture. To simultaneously recognize Staphylococci genus, S. aureus and methicillin resistance, three sets of six primers for 16S rRNA, femA and mecA were designed, respectively. The performance of LAMP assay was affirmed using VITEK system for the phenotypic methods of identification and for oxacillin and cefoxitin antimicrobial susceptibility. The optimal condition for LAMP assay was obtained under 64꼦 for 50 min. The detection limit was determined to be of 20 copies and CFU/reaction (104 CFU/mL). For clinical application of comparison with phenotypic methods, the sensitivity and specificity of the LAMP with femA gene for detecting S. aureus was 95.31% and 100%, respectively. The sensitivity and specificity of the LAMP with mecA gene for detecting methicillin resistance was 98.46% and 100%, respectively. The multiplex LAMP assay with femA and mecA gene successfully detected all of MRSA (38 isolates) isolates from 103 Staphylococci in blood cultures. The LAMP assay developed in this study is sensitive, specific, and of excellent agreement with the phenotypic methods.
Keywords : Blood culture, Methicillin-resistant Staphylococcus aureus, 16S rRNA, femA, mecA, Loop-mediated isothermal amplification