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Employment of 16S rDNA gene sequencing techniques for improved identification of difficult-to-identify bacterial veterinary pathogens |
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| Authors: | John E Moore Yasunori Maeda Jiru Xu B Cherie Millar Peter H Herold V M J Browne-Lauwers Colin E Goldsmith Anne Loughrey Paul J Rooney J Stuart Elborn Motoo Matsuda |
| Institution: | 1. Northern Ireland Public Health Laboratory, Department of Bacteriology, Belfast City Hospital, Lisburn Road, Belfast, BT9 7AD, Northern Ireland, UK 2. Department of Pathogenic Biology, Xian Jiatong University, Xi’an, Shaanxi, The People’s Republic of China 3. Gortlands Veterinary Clinic, 162 Gilnahirk Road, Belfast, BT5 7QQ, Northern Ireland, UK 4. Northern Ireland Regional Adult Cystic Fibrosis Centre, Level 8, Belfast City Hospital, Belfast, BT9 7AB, Northern Ireland, UK 5. Department of Respiratory Medicine, School of Medicine and Dentistry, Queen’s University, Level 8, Belfast City Hospital, Belfast, BT9 7AB, Northern Ireland, UK 6. Laboratory of Molecular Biology, School of Environmental Health Sciences, Azabu University, Fuchinobe 1-17-71, Sagamihara, 229-8501, Japan |
| Abstract: | To employ 16S rDNA PCR and automated sequencing techniques to identify a collection of bacterial veterinary pathogens from avian, equine, canine and ovine sources, that have proven difficult to identify, employing conventional cultural techniques. Universal or “broad-range” eubacterial PCR was performed on a collection of 46 difficult-to-identify bacterial isolates originating from clinical veterinary specimens. 16S rDNA PCR was performed using two sets of universal primers to successfully generate a composite amplicon of 1,068 bp, which was sequenced to obtain each isolate’s identity. Sequence analysis was able to identify all isolates examined with relative ease. Where the use of molecular identification methods is justified, such as in outbreak control or bioterrorism in animal health, employment of partial 16S rDNA PCR and sequencing employing universal or “broad-range” 16S rDNA, provides a valuable and reliable method of identification of such pathogens. |
| Keywords: | Pseudomonas Taylorella Bacteroides Campylobacter UPTC PCR 16S rRNA Molecular diagnosis Pyoderma |
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