Peer-reviewed veterinary case report
Validation of a high-throughput real-time polymerase chain reaction assay for the detection of capripoxviral DNA.
- Journal:
- Journal of virological methods
- Year:
- 2012
- Authors:
- Stubbs, Samuel et al.
- Affiliation:
- Institute for Animal Health · United Kingdom
Plain-English summary
This study focused on improving a test to quickly identify capripoxviruses, which cause serious diseases in cattle, sheep, and goats, especially in Africa and Asia. Researchers compared two existing testing methods and found that their new approach was more effective at detecting the virus without mistakenly identifying similar viruses. They tested samples from animals known to be infected and confirmed that their method could accurately find the virus even when using automated equipment for sample preparation. This new testing method is reliable and can be used in labs to help diagnose capripoxvirus infections more efficiently. Overall, the new test works well and is a valuable tool for detecting these viruses.
Abstract
Capripoxviruses, which are endemic in much of Africa and Asia, are the aetiological agents of economically devastating poxviral diseases in cattle, sheep and goats. The aim of this study was to validate a high-throughput real-time PCR assay for routine diagnostic use in a capripoxvirus reference laboratory. The performance of two previously published real-time PCR methods were compared using commercially available reagents including the amplification kits recommended in the original publication. Furthermore, both manual and robotic extraction methods used to prepare template nucleic acid were evaluated using samples collected from experimentally infected animals. The optimised assay had an analytical sensitivity of at least 63 target DNA copies per reaction, displayed a greater diagnostic sensitivity compared to conventional gel-based PCR, detected capripoxviruses isolated from outbreaks around the world and did not amplify DNA from related viruses in the genera Orthopoxvirus or Parapoxvirus. The high-throughput robotic DNA extraction procedure did not adversely affect the sensitivity of the assay compared to manual preparation of PCR templates. This laboratory-based assay provides a rapid and robust method to detect capripoxviruses following suspicion of disease in endemic or disease-free countries.
Find similar cases for your pet
PetCaseFinder finds other peer-reviewed reports of pets with the same symptoms, plus a plain-English summary of what was tried across them.
Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/22138682/