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Published (Version of Record)CC BY V4.0, Open Access
Journal article
Assessment of a rtPCR for the detection of virulent and benign Dichelobacter nodosus, the causative agent of ovine footrot, in Australia
BMC veterinary research, Vol.14(1), 252
2018
PMCID: PMC6114850
PMID: 30157841
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.
Details
- Title
- Assessment of a rtPCR for the detection of virulent and benign Dichelobacter nodosus, the causative agent of ovine footrot, in Australia
- Authors/Creators
- Nickala Best - La Trobe UniversityLucas Zanandrez - Universidade Federal de Minas GeraisJacek Gwozdz - Government of VictoriaEckard Klien - DAFWA Diagnostics and Laboratory Services, Biosecurity and Regulation, Department of Agriculture and Food, South Perth, Western Australia.Nicky Buller - DAFWA Diagnostics and Laboratory Services, Biosecurity and Regulation, Department of Agriculture and Food, South Perth, Western Australia.Robert Suter - Department of Economic Development Jobs Transport and ResourcesGrant Rawlin - Government of VictoriaTravis Beddoe - AgriBio
- Publication Details
- BMC veterinary research, Vol.14(1), 252
- Publisher
- Springer Nature
- Number of pages
- 12
- Grant note
- Victorian Sheep and Goat Compensation fund
- Identifiers
- 991005579393907891
- Copyright
- © The Author(s) 2018
- Murdoch Affiliation
- Harry Butler Institute; Centre for Sustainable Aquatic Ecosystems
- Language
- English
- Resource Type
- Journal article
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- Citation topics
- 3 Agriculture, Environment & Ecology
- 3.51 Dairy & Animal Sciences
- 3.51.799 Farm Animal Welfare
- Web Of Science research areas
- Veterinary Sciences
- ESI research areas
- Plant & Animal Science