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- Title
Assessment of a rtPCR for the detection of virulent and benign Dichelobacter nodosus, the causative agent of ovine footrot, in Australia.
- Authors
Best, Nickala; Zanandrez, Lucas; Gwozdz, Jacek; Klien, Eckard; Buller, Nicky; Suter, Robert; Rawlin, Grant; Beddoe, Travis
- Abstract
Background: Ovine footrot is a highly contagious bacterial disease of sheep, costing the Australian sheep industry millions of dollars annually. Dichelobacter nodosus, the causative agent of footrot, is a gram-negative anaerobe classed into virulent and benign strains as determined by thermostability of their respective protesases. Current methods for detection of D. nodosus are difficult and time-consuming, however new molecular techniques capable of rapidly detecting and typing D. nodosus have been reported. Results: A competitive real-time PCR (rtPCR) method, based on the ability to detect a 2 nucleotide difference in the aprV2 (virulent) and aprB2 (benign) extracellular protease gene has been tested on Australian samples for determining detection rates, along with clinically relevant cut-off values and performance in comparison to the traditional culturing methods. The rtPCR assay was found to have a specificity of 98.3% for virulent and 98.7% for benign detection from samples collected. Sheep with clinical signs of footrot showed a detection rate for virulent strains of 81.1% and for benign strains of 18.9%. A cut-off value of a Ct of 35 was found to be the most appropriate for use in Victoria for detection of sheep carrying virulent D. nodosus. Conclusions: In summary, the rtPCR assay is significantly more capable of detecting D. nodosus than culturing, while there is no significant difference seen in virotyping between the two methods.
- Subjects
AUSTRALIA; POLYMERASE chain reaction; FOOTROT in sheep; GRAM-negative anaerobic bacteria; PROTEOLYTIC enzymes
- Publication
BMC Veterinary Research, 2018, Vol 14, Issue 1, pN.PAG
- ISSN
1746-6148
- Publication type
Article
- DOI
10.1186/s12917-018-1575-0