Peer-reviewed veterinary case report
Development of a RT-PCR ELISA for simultaneous detection of BVDV-1, BVDV-2 and BDV in ruminants and its evaluation on clinical samples.
- Journal:
- Journal of virological methods
- Year:
- 2015
- Authors:
- Dubey, Pooja et al.
- Affiliation:
- National Institute of High Security Animal Diseases (formerly HSADL) · India
Plain-English summary
This study focused on creating a new test called RT-PCR ELISA to detect certain viruses in cattle, sheep, and goats. The researchers found that this test could reliably identify the viruses at very low levels, and it performed well when compared to other testing methods. When they tested samples from infected animals, the RT-PCR ELISA showed a high success rate in correctly identifying the viruses, with 95.9% sensitivity (how well it detects true positives) and 98.6% specificity (how well it avoids false positives). The results suggest that this new test could be a quick and cost-effective way to diagnose these viruses in farm animals. Overall, the treatment worked well, showing that the RT-PCR ELISA is a promising option for detecting these viruses.
Abstract
The aim of this study was to develop a reverse transcription polymerase chain reaction ELISA (RT-PCR ELISA) for detection of ruminant pestiviruses and to evaluate its diagnostic performance on clinical samples obtained from cattle, sheep and goats. Optimization was carried out by serial dilution of home-made digoxygenin-labelled RT-PCR product standards obtained from pestivirus isolates and pestivirus infected animals. The detection limit of the assay was 10TCID50/ml, similar to virus isolation and real-time RT-PCR but 10-fold higher than RT-PCR. The assay had high analytical specificity along with a good reproducibility. When the assay was evaluated on the samples obtained from animals infected experimentally with BVDV and from the field using virus isolation as standard, it showed a high diagnostic sensitivity (95.9%) and specificity (98.6%) and there was strong agreement (97.5% concordance) between the two tests. However, it displayed an increased diagnostic specificity and sensitivity over RT-PCR. Additionally, when a few samples (n=26) were tested by RT-PCR ELISA and real-time RT-PCR, 100% concordance was obtained between them. Our results showed that RT-PCR ELISA can be a rapid, cost effective and alternative molecular diagnostic test for simultaneous detection of BVDV-1, BVDV-2 and BDV in ruminants in ordinary laboratory settings.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/25486086/