title: Rapid detection and differentiation of important Campylobacter spp. in poultry samples by dot blot and PCR.
abstract: The detection of Campylobacter, the most commonly reported cause of foodborne gastroenteritis in the European Union, is very important for human health. The most commonly recognised risk factor for infection is the handling and/or consumption of undercooked poultry meat. The methods typically applied to evaluate the presence/absence of Campylobacter in food samples are direct plating and/or enrichment culture based on the Horizontal Method for Detection and Enumeration of Campylobacter spp. (ISO 10272-1B: 2006) and PCR. Molecular methods also allow for the detection of cells that are viable but cannot be cultivated on agar media and that decrease the time required for species identification. The current study proposes the use of two molecular methods for species identification: dot blot and PCR. The dot blot method had a sensitivity of 25 ng for detection of DNA extracted from a pure culture using a digoxigenin-labelled probe for hybridisation; the target DNA was extracted from the enrichment broth at 24 h. PCR was performed using a pair of sensitive and specific primers for the detection of Campylobacter jejuni and Campylobacter coli after 24 h of enrichment in Preston broth. The initial samples were contaminated by 5 × 10 C. jejuni cells/g and 1.5 × 10(2)C. coli cells/g, thus the number of cells present in the enrichment broth at 0 h was 1 or 3 cell/g, respectively.
authors.affiliation: Department of Food Science, University of Udine, Via Sondrio 2/A, 33100 Udine, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Fontanot
authors.affiliation: Department of Food Science, University of Udine, Via Sondrio 2/A, 33100 Udine, Italy.
authors.forename: Lucilla
authors.initials: L
authors.lastname: Iacumin
authors.affiliation: Department of Food Science, University of Udine, Via Sondrio 2/A, 33100 Udine, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Cecchini
authors.affiliation: Department of Food Science, University of Udine, Via Sondrio 2/A, 33100 Udine, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Comi
authors.affiliation: Department of Food Science, University of Udine, Via Sondrio 2/A, 33100 Udine, Italy. Electronic address: marisa.manzano@uniud.it.
authors.forename: Marisa
authors.initials: M
authors.lastname: Manzano
mesh_terms: D000818:Animals; D015373:Bacterial Typing Techniques; D002167:Campylobacter / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002645:Chickens; D005506:Food Contamination / Q000032:analysis*; D008460:Meat / Q000032:analysis*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014422:Turkeys
keywords: 16S and ITS sequences; Campylobacter; Dot blot; PCR; Poultry meat contamination