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KMID : 1144820190250010075
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대한의생명과학회지
2019 Volume.25 No. 1 p.75 ~ p.82
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Application of Loop-Mediated Isothermal Amplification (LAMP) Assay to Rapid Detection of Methicillin-Resistant Staphylococcus aureus from Blood Cultures
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Baek Yun-Hee
Jo Mi-Young
Song Min-Suk
Hong Seung-Bok
Shin Kyeong-Seob
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Abstract
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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 64℃ 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.
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KEYWORD
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Blood culture, Methicillin-resistant Staphylococcus aureus, 16S rRNA, femA, mecA, Loop-mediated isothermal amplification
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