Sensitive and Specific TaqMan Real-Time PCR Assay for Beak and Feather Disease Virus in Psittacine Birds
Year: 2025
Type: article
Abstract: family. BFDV is globally distributed and poses a major threat to susceptible avian populations. This underscores the urgent need for rapid and reliable molecular detection methods to monitor and control the spread of the virus. Quantitative PCR assays offer several advantages, including high sensitivity, specificity, and the ability to quantify viral load. Here, we report the development and validation of a quantitative real-time PCR assay using a TaqMan probe targeting the replicase gene to detect BFDV in psittacine blood samples. The assay demonstrates high sensitivity, specificity, and suitability for routine diagnostic use with an LOD and an LOQ of 10 and 30 plasmid copies, respectively, determined using a recombinant plasmid containing a BFDV genomic fragment. The assay achieved 98.8% sensitivity and 100% specificity. The method also demonstrated strong repeatability and reproducibility, with intra- and inter-assay coefficients of variation of 4.14% and 4.44%, respectively. Both values are well below the acceptance threshold. Overall, this TaqMan-based real-time PCR assay is a reliable, sensitive, and efficient diagnostic tool for detecting BFDV in psittacine samples.
Source: Veterinary Sciences
Authors: Bruno Fernandes, Teresa Fagulha, Sílvia Carla Barros, Fernanda Ramos, Tiago Luís, Ana Duarte, Margarida Dias Duarte, Ana Margarida Henriques
Language: English
Subfield: Animal Science and Zoology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,534
OpenAlex ID: W4416944768