Pasteurella multocida detection by 5′ Taq nuclease assay: A new tool for use in diagnosing fowl cholera

• Published in: Journal of microbiological methods 2007-05, Vol.69 (2), p.376-380
• Main Authors: Corney, B.G., Diallo, I.S., Wright, L.L., Hewitson, G.R., Jong, A.J. De, Burrell, P.C., Duffy, P.F., Stephens, C.P., Rodwell, B.J., Boyle, D.B., Blackall, P.J.
• Format: Article
• Language: English
• Subjects: 5′ Taq nuclease assay; Animals; Bacteriological methods and techniques used in bacteriology; Bacteriological Techniques - methods; Bacteriology; Base Sequence; Biological and medical sciences; Cattle; DNA Primers; DNA Restriction Enzymes - chemistry; DNA Restriction Enzymes - metabolism; Fowl cholera; Fundamental and applied biological sciences. Psychology; Microbiology; Minor groove binder; Molecular Sequence Data; Pasteurella; Pasteurella Infections - diagnosis; Pasteurella Infections - microbiology; Pasteurella Infections - veterinary; Pasteurella multocida; Pasteurella multocida - genetics; Pasteurella multocida - isolation & purification; Pasteurellaceae; PCR; Poultry; Poultry Diseases - diagnosis; Poultry Diseases - microbiology; RNA, Ribosomal, 16S - metabolism; Swine
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2007.01.014

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.