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Updated: Jan 6, 2026
Development and Validation of a Quantitative PCR Method for Equid Herpesvirus-2 Diagnostics in Respiratory Fluids
Published on: March 17, 2016
Development of a TaqMan based real-time PCR assay for the detection of bovine ephemeral fever virus
Shivam Solanki1, M Manu2, Adarsh Mishra1
1Department of Microbial and Environmental Biotechnology, College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, 141004, India.
Background:
Bovine ephemeral fever (BEF) is a significant disease in nations such as India, where there is a large bovine population and environmental conditions which foster the proliferation of blood-feeding insects. Effective control and prevention of the disease necessitate early and precise detection of bovine ephemeral fever virus (BEFV), the causative agent of BEF. In the present study, a TaqMan real time PCR assay with high sensitivity and specificity was developed for accurate detection of BEFV.
Methods And Results:
The glycoprotein (G) gene of BEFV was targeted for the assay development. A positive construct of the G gene was developed by identifying the conserved regions through the alignment of the available G gene sequences from GenBank. The primers and probes had been designed in-house and synthesised commercially. The TaqMan probe-based assay was standardized using the gene construct as a positive control. The developed assay demonstrated an exceptional sensitivity for the detection and quantification of viral genomes, capable of identifying as few as 24 copies of the genome with a R² value of 0.9998 by standard curve analysis. The developed assay demonstrated a sensitivity that was 100 times greater than the conventional PCR method and showed remarkable efficiency for the BEFV detection using field blood samples from both suspected and healthy animals.
Conclusion:
The developed assay showed high sensitivity and specificity in clinical applicability analysis. This real-time PCR-based assay would offer a reliable and adaptable diagnostic tool for the rapid and precise identification of BEFV in disease diagnosis, research, and field surveillance applications.
