title: Development and evaluation of TaqMan real-time PCR assay for detection of beak and feather disease virus.
abstract: Psittacine beak and feather disease (PBFD) is one of the most significant viral diseases in psittacine birds. The aim of the presented study was to develop a highly specific and sensitive TaqMan real-time PCR assay for universal detection of beak and feather disease virus (BFDV). Primers and a hydrolysis probe were selected on the highly conserved regions belonging to the ORF1 of the BFDV genome which were identified by aligning 814 genomic sequences downloaded from the GenBank database. The evaluation of the reaction parameters suggested a reaction efficiency of 97.1%, with consistent detection of 101virus copies/μl of nucleic acid extract. The low values of standard deviation and coefficient of variation indicate a high degree of reproducibility and repeatability. The diagnostic applicability of the assay was proven on 36 BFDV positive and 107 negative specimens of psittacine origin representing 28 species. The assay showed a 100% ability to detect distinct genetic variants of the virus. Our data suggest that the presented TaqMan real-time PCR represents a specific, sensitive and reliable assay facilitating the molecular detection of BFDV.
authors.affiliation: State Veterinary Institute Prague, Prague, Czechia; University of Veterinary and Pharmaceutical Sciences, Department of Biology and Wildlife Diseases, Brno, Czechia. Electronic address: len.cernikova@gmail.com.
authors.forename: Lenka
authors.initials: L
authors.lastname: Černíková
authors.affiliation: State Veterinary Institute Prague, Prague, Czechia; University of Veterinary and Pharmaceutical Sciences, Department of Biology and Wildlife Diseases, Brno, Czechia.
authors.forename: Eliška
authors.initials: E
authors.lastname: Vitásková
authors.affiliation: State Veterinary Institute Prague, Prague, Czechia. Electronic address: alexandernagy17@hotmail.com.
authors.forename: Alexander
authors.initials: A
authors.lastname: Nagy
mesh_terms: D000818:Animals; D001715:Bird Diseases / Q000175:diagnosis* / Q000821:virology*; D001717:Birds; D018173:Circoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D017925:Circovirus / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D017931:DNA Primers; D015345:Oligonucleotide Probes
keywords: Beak and feather disease virus (BFDV); Real-time PCR