title: Development of 23 individual TaqMan® real-time PCR assays for identifying common foodborne pathogens using a single set of amplification conditions.
abstract: Most of the acute intestinal diseases are caused by foodborne pathogens with infants and elderly people being at major risk. The aim of this study was to develop a procedure to simultaneously detect 20 foodborne pathogens in complex alimentary matrices such as milk, cheese and meat. The list of targets include, among the others, Listeria spp., Salmonella spp., Shigella spp., Escherichia coli spp., Campylobacter spp., Clostridium spp. and Staphylococcus aureus. The accuracy of detection was determined by using ATCC strains as positive and negative controls. The achieved sensitivity of each of assays was 1 pg of genomic DNA, which was equivalent to ∼1 cfu. The working ranges of the TaqMan(®) Real-time PCR assays, when used quantitatively on cheese and meat samples inoculated with serial dilution of Listeria spp., Listeria monocytogenes, S. aureus, Salmonella enterica, Shigella boydii, E. coli O157:H7, Bacillus cereus, Campylobacter coli, Yersinia enterocolitica, Enterobacter sakazakii and Pseudomonas aeruginosa was 10(8) cfu/g to 10(4) cfu/g. No matrix interferences were observed.
authors.affiliation: Institute of Agricultural Biology and Biotechnology, National Research Council, via Einstein, 26900 Lodi, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Cremonesi
authors.affiliation: Department of Veterinary Science and Public Health, via Celoria 10, 20133 Milan, Italy.
authors.forename: Laura Francesca
authors.initials: LF
authors.lastname: Pisani
authors.affiliation: Department of Veterinary Science and Public Health, via Celoria 10, 20133 Milan, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Lecchi
authors.affiliation: Department of Veterinary Science and Public Health, via Celoria 10, 20133 Milan, Italy.
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Ceciliani
authors.affiliation: Department of Veterinary Science and Public Health, via Celoria 10, 20133 Milan, Italy.
authors.forename: Pieranna
authors.initials: P
authors.lastname: Martino
authors.affiliation: Universitat Autonoma de Barcelona, Campus de la UAB, Bellaterra, Spain.
authors.forename: Armand Sanchez
authors.initials: AS
authors.lastname: Bonastre
authors.affiliation: Department of Food Sciences and Hygiene, Institute of Veterinary Medicine and Animal Science, Estonian University of Life Sciences, Kreutzwaldi 62, Tartu, Estonia.
authors.forename: Avo
authors.initials: A
authors.lastname: Karus
authors.affiliation: Department of Health, Animal Science and Food Safety, via Celoria 10, 20133 Milan, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Balzaretti
authors.affiliation: Institute of Agricultural Biology and Biotechnology, National Research Council, via Einstein, 26900 Lodi, Italy. Electronic address: casti@ibba.cnr.it.
authors.forename: Bianca
authors.initials: B
authors.lastname: Castiglioni
mesh_terms: D000818:Animals; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002417:Cattle; D002611:Cheese / Q000382:microbiology*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D008461:Meat Products / Q000382:microbiology*; D008892:Milk / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D013552:Swine; D019914:Taq Polymerase / Q000737:chemistry
chemical_list: D019914:Taq Polymerase
keywords: Cheese; Foodborne pathogens; Meat; TaqMan(®) real-time quantitative PCR