title: A multiplex PCR assay for simultaneous detection of Escherichia coli O157:H7, Bacillus cereus, Vibrio parahaemolyticus, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus in Korean ready-to-eat food.
abstract: A multiplex polymerase chain reaction (PCR) assay was developed for simultaneous detection of Escherichia coli O157:H7, Bacillus cereus, Vibrio parahaemolyticus, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus in various Korean ready-to-eat foods. The six specific primer pairs for multiplex PCR were selected based on the O157 antigen (rfbE) gene of E. coli O157:H7, the DNA gyrase subunit B (gyrB) gene of B. cereus, the toxin regulatory protein (toxR) gene of V. parahaemolyticus, the invasion protein A (invA) gene of Salmonella spp., the hemolysin (hly) gene of L. monocytogenes, and the thermonuclease (nuc) gene of S. aureus. The 16S rRNA gene was targeted as an internal control gene in the presence of bacterial DNA. The specificity and sensitivity assays for multiplex primer pairs were investigated by testing different strains. When this multiplex PCR assay was applied to evaluate the validity of detecting six foodborne pathogens in artificially inoculated several ready-to-eat food samples, the assay was able to specifically simultaneously detect as few as 1 colony-forming unit/mL of each pathogen after enrichment for 12 h. Their presence in naturally contaminated samples also indicates that the developed multiplex PCR assay is an effective and informative supplement for practical use.
authors.affiliation: 1 Food Safety Research Group, Korea Food Research Institute , Gyeonggi, Korea.
authors.forename: Nari
authors.initials: N
authors.lastname: Lee
authors.forename: Kyung Yoon
authors.initials: KY
authors.lastname: Kwon
authors.forename: Su Kyung
authors.initials: SK
authors.lastname: Oh
authors.forename: Hyun-Joo
authors.initials: HJ
authors.lastname: Chang
authors.forename: Hyang Sook
authors.initials: HS
authors.lastname: Chun
authors.forename: Sung-Wook
authors.initials: SW
authors.lastname: Choi
mesh_terms: D001409:Bacillus cereus / Q000302:isolation & purification; D015169:Colony Count, Microbial; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D019453:Escherichia coli O157 / Q000302:isolation & purification; D057140:Fast Foods / Q000382:microbiology*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D008089:Listeria monocytogenes / Q000302:isolation & purification; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D056910:Republic of Korea; D012475:Salmonella / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000302:isolation & purification; D014736:Vibrio parahaemolyticus / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S