Novel multiplex RT-PCR/RFLP diagnostic test to differentiate low- from high-pathogenic strains and to detect reassortant infectious bursal disease virus.
Avian Dis
; 55(3): 368-74, 2011 Sep.
Article
em En
| MEDLINE
| ID: mdl-22017032
ABSTRACT
Three types of infectious bursal disease virus (IBDV) strains are currently circulating worldwide the low-pathogenic classic and variant strains and the high-pathogenic very virulent strains. There are also natural reassortant viruses that combine genomic segments A and B from different strains and exhibit particular pathogenic characteristics. Detection and characterization of the different IBDVs is extremely critical for improving disease control and performing epidemiologic studies. Here, we present a novel detection and genotyping method based on the simultaneous characterization of both IBDV genomic segments followed by a simple restriction fragment length polymorphism (RFLP) assay. This single restriction enzyme, multiplex reverse transcriptase-PCR/RFLP diagnostic test not only distinguished typical high-pathogenic from low-pathogenic strains but also detected natural reassortant IBDV. The test was based on the detection of single nucleotide polymorphisms (SNP), in both segments, which were strongly linked to the pathogenic phenotype. These SNPs are embedded in highly conserved genomic regions and can be identified with TfiI endonuclease. The application of this methodology in field samples confirmed that the assay is fast, specific, and may be easily adopted by any molecular diagnostic laboratory as an economical and routine method.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Doenças das Aves Domésticas
/
Bolsa de Fabricius
/
Infecções por Birnaviridae
/
Vírus da Doença Infecciosa da Bursa
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Animals
Idioma:
En
Revista:
Avian Dis
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Uruguai