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KMID : 1134820100390040595
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Journal of the Korean Society of Food Science and Nutrition
2010 Volume.39 No. 4 p.595 ~ p.601
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Simultaneous Detection of Staphylococcus aureus, Salmonella enterica subsp., Vibrio parahaemolyticus by Multiplex Polymerase Chain Reaction
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Jeong Yoo-Seok
Jung Hee-Kyoung
Seo Hwa-Jung
Hong Joo-Heon
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Abstract
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This study was conducted to detect and identify Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella enterica subsp. using simultaneous multiplex polymerase chain reaction (multiplex PCR) assay. 23S rRNA partial gene (S. aureus), tox R gene (V. parahaemolyticus), and inv A gene (S. enterica subsp.) as diagnostic marker gene were suggested, and their amplicon sizes were 482 bp, 368 bp, and 284 bp, respectively. Non specific amplicons by STA-5F/STA-5R primer, ToxR-F/ToxR-R primer, and 139/141 primer were not observed in genomic DNA of pathogen bacteria as Bacillus cereus, Listeria monocytogenes, Escherichia coli, Proteus vulgaris, Streptococcus pyogenes, Candida albicans, and Shigella sonnei. The extracted crude DNA of targeted bacteria was detected as PCR template successfully. The detection limits were 105~104 CFU/mL and 10 pg of purified genomic DNA of S. aureus, V. parahaemolyticus, and S. enterica subsp. by using simultaneous multiplex PCR.
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KEYWORD
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Staphylococcus aureus, Vibrio parahaemolyticus, Salmonella enterica subsp., multiplex PCR
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