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KMID : 0361319940260010269
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Korean Journal of Medical Technologists
1994 Volume.26 No. 1 p.269 ~ p.272
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Rapid Detection of the mecA Gene in Methicillin-Resistant Staphylococcus aureus
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Park Jeong-Jun
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Abstract
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In order to identify methicillin-resistant Staplyloccus aureus from clinical sources with ease and reliability, enzymatic detection of polymerase chain reaction(ED-PCR) was applied. ED-PCR is based on the capture of amplified products via biotin-streptavidn affinity and the detection of and incorporated hapten in amplified products with an enzyme-linked antibody. In order to identify methicillin-resistant staphyloccus aureus, a 150-bp fragment of the mecA gene was targeted for ED-RCR. After PCR was performed with a pair of biotin and dinitrophenol 5’-labeled primers, the reaction mixture was applied to a microtiter well precoated with streptavidin. Thereafter, bound PCR products were detected colorimetrically with alkaline phosphatase conjugated anti-dinitrophenol antibody. The extraction of DNA form Staphylococcus aureus for PCR was simplified so that it could be performed within one tube. The total assay, including PCR, took less than 3h.
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KEYWORD
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Methicillin-resistant Staplylococcus aureus, ED-PCR
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