Detection and differentiation of Campylobacter jejuni and Campylobacter coli in broiler chicken samples using a PCR/DNA probe membrane based colorimetric detection assay.
Campylobacter enteritis in humans has been linked to consumption of poultry meat. Surveys show that 30-100% of poultry harbour Campylobacter as normal flora of the digestive tract which indicates a need to identify prevalent organism types in flocks and trace their epidemiology. In this study we describe a Campylobacter genus specific polymerase chain reaction (PCR) assay, amplifying the 16 S-23 S rRNA intergenic spacer region with an internal Campylobacter genus specific DNA probe and species specific probes for Campylobacter jejuni and Campylobacter coli designed for confirmation of the amplified PCR products by Southern blot and colorimetric reverse hybridization assays. The specificity of this assay was established by testing a range of food pathogens. Broiler chicken samples were tested following presumptive positive identification by the Malthus System V analyser (Malthus Instruments, UK). The combined PCR and colorimetric reverse hybridization assay is easy to perform and faster than conventional methods for confirmation and identification of Campylobacter species.
O'Sullivan
N A
NA
DNA Diagnostics, National Diagnostics Centre, Galway, Ireland.
Fallon
R
R
Carroll
C
C
Smith
T
T
Maher
M
M
D000818:Animals; D015139:Blotting, Southern; D017000:Campylobacter coli / Q000235:genetics* / Q000302:isolation & purification; D016123:Campylobacter jejuni / Q000235:genetics* / Q000302:isolation & purification; D002645:Chickens / Q000382:microbiology; D002863:Chromogenic Compounds; D003124:Colorimetry; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry; D005516:Food Microbiology; D007211:Indoles; D008460:Meat / Q000382:microbiology*; D008969:Molecular Sequence Data; D009580:Nitroblue Tetrazolium; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D011201:Poultry Diseases / Q000382:microbiology*; D012335:RNA, Ribosomal / Q000737:chemistry; D012680:Sensitivity and Specificity
D002863:Chromogenic Compounds; D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; D007211:Indoles; D012335:RNA, Ribosomal; D009580:Nitroblue Tetrazolium; C035455:5-bromo-4-chloro-3-indoxyl phosphate