A universal protocol for PCR detection of 13 species of foodborne pathogens in foods

Abstract

was developed. The protocol used a universal culture medium and the same PCR conditions with 13 sets of specific primers. The 13 species of foodborne pathogens examined were Escherichia coli, E. coli-ETEC, E. coli-O157:H7, Shigella spp., Salmonella spp., Yersinia enterocolitica, Y. pseudotuberculosis, Vibrio cholerae, V. parahaemolyticus, V. vulnificus, Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus. No interference was observed using the PCR assay when food sample was artificially inoculated with each individual bacterial species. Twelve different seafood samples and two soft cheese samples without artificial inoculation were examined by this protocol. Vibrio vulnificus, Salmonella spp., E. coli, Listeria monocytogenes and Bacillus cereus were detected in some foods. Internal probe hybridization and nested PCR procedures were used to confirm the above findings.

Screening of lactic acid bacteria for bacteriocins by microbiological and PCR methods

“…In teaching, the principle of PCR has facilitated student understanding of DNA structure and replication. PCR has previously been shown to be useful in student laboratory practice, particularly to detect genetically modified organisms (GMOs) [4][5][6] and to identify and characterize pathogenic microorganisms [7,8].…”

Reverse Transcription-Multiplex PCR Assay for Simultaneous Detection of Escherichia coli O157:H7, Vibrio cholerae O1, and Salmonella Typhi

“…Molecular techniques such as PCR have been invaluable tools for the detection of pathogens (12 ). When multiple target genes need to be amplified, multiplex PCR (MPCR) can be performed and may provide a simple and sensitive tool for the simultaneous detection of multiple pathogenic bacteria (13 ).…”

“…PCR, however, does not provide information related to cell viability because it cannot distinguish the DNA molecules from live and dead cells (14,15 ). To improve discrimination power, a preenrichment step is typically performed to allow viable cells to grow and multiply (12 ). However, the preenrichment process takes 6 -48 h to complete, which becomes the bottleneck for the subsequent PCR (16 ).…”

Prevalence of Staphylococcus aureus and staphylococcal enterotoxins in raw pork and uncooked smoked ham—a comparison of classical culturing detection and RFLP-PCR

“…1995;Wang et al, 1997;Pimbley and Patel, 1998 . Methods have been described for the identification Ž of S. aureus Johnson et al, 1991;Brakstad et al, .…”