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KMID : 1059520130570010081
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Journal of the Korean Chemical Society
2013 Volume.57 No. 1 p.81 ~ p.87
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Detection for Methicillin Resistant Staphylococcus aureus in Using Bio-Chip Based Loop Mediated Isothermal Amplification Assay
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Cho Min-Ho
Jang Won-Choeul
Choi Jae-Gu
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Abstract
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Staphylococcus aureus is the most important pathogen in nosocomial infections, including bloodstream infections. Prompt identification of S. aureus from blood cultures and detection of methicillin resistance are essential in cases of suspected sepsis. We have studied a new method for the sequence-specific visual detection of minute amounts of nucleic acids using intercalating reaction by addition of SYBR Green to amplicons of LAMP, and it’s a unique gene amplification method in which DNA can be isothermally amplified using only one enzyme. Staphylococcus-LAMP, which targets the spa gene, encoding S.aureus-specific protein A, and the mecA gene, encoding penicillin-binding protein-2’ for methicillin resistance, detected MRSA and MRSE. In this study, by using LAMP assay, I detected for Staphylococcus aureus and Staphylococcus epidermidis concentration in the clinical sample. The detection of Staphylococcus aureus and Staphylococcus epidermidis was tested by using serial 10-fold dilutions standard solution. I have accurate detected the limit of detection, sensitity, specificity and reproducibility of the assay. The Bio-chip based LAMP assay allowed easy, rapid, accurate and sensitive detection of infection with Staphylococcus and especially applicable in a resource-limited situation.
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KEYWORD
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MRSA (methicillin resistant Staphylococcus aureus), LAMP (loop mediated isothermal amplification), SYBR Green I, Bio-chip
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