title: Pasteurella multocida detection by 5' Taq nuclease assay: a new tool for use in diagnosing fowl cholera.
abstract: A 5' Taq nuclease assay utilising minor groove binder technology and targeting the 16S rRNA gene was designed to detect Pasteurella multocida (the causative agent of fowl cholera) in swabs collected from poultry. The assay was first evaluated using pure cultures. The assay correctly identified four P. multocida taxonomic type strains, 18 P. multocida serovar reference strains and 40 Australian field isolates (17 from poultry, 11 from pigs and 12 from cattle). Representatives of nine other Pasteurella species, 26 other bacterial species (18 being members of the family Pasteurellaceae) and four poultry virus isolates did not react in the assay. The assay detected a minimum of approximately 10 cfu of P. multocida per reaction. Of 79 poultry swabs submitted to the laboratory for routine bacteriological culture, 17 were positive in the 5' Taq nuclease assay, but only 10 were positive by culture. The other 62 swabs were negative for P. multocida by both 5' Taq nuclease assay and culture. The assay is suitable for use in diagnosing fowl cholera, is more rapid than bacteriological culture, and may also have application in diagnosing P. multocida infections in cattle and pigs.
authors.affiliation: Animal Research Institute, Department of Primary Industries and Fisheries, Yeerongpilly, Queensland 4105, Australia. bruce.corney@dpi.qld.gov.au
authors.forename: B G
authors.initials: BG
authors.lastname: Corney
authors.forename: I S
authors.initials: IS
authors.lastname: Diallo
authors.forename: L L
authors.initials: LL
authors.lastname: Wright
authors.forename: G R
authors.initials: GR
authors.lastname: Hewitson
authors.forename: A J
authors.initials: AJ
authors.lastname: De Jong
authors.forename: P C
authors.initials: PC
authors.lastname: Burrell
authors.forename: P F
authors.initials: PF
authors.lastname: Duffy
authors.forename: C P
authors.initials: CP
authors.lastname: Stephens
authors.forename: B J
authors.initials: BJ
authors.lastname: Rodwell
authors.forename: D B
authors.initials: DB
authors.lastname: Boyle
authors.forename: P J
authors.initials: PJ
authors.lastname: Blackall
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods; D001483:Base Sequence; D002417:Cattle; D017931:DNA Primers; D004262:DNA Restriction Enzymes / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D010326:Pasteurella Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D016979:Pasteurella multocida / Q000235:genetics / Q000302:isolation & purification*; D011200:Poultry; D011201:Poultry Diseases / Q000175:diagnosis / Q000382:microbiology*; D012336:RNA, Ribosomal, 16S / Q000378:metabolism; D013552:Swine
chemical_list: D017931:DNA Primers; D012336:RNA, Ribosomal, 16S; D004262:DNA Restriction Enzymes