Differentiation of infectious bursal disease virus strains using real-time RT-PCR and high resolution melt curve analysis.
J Virol Methods
; 171(1): 264-71, 2011 Jan.
Article
em En
| MEDLINE
| ID: mdl-21111004
ABSTRACT
Differentiation of infectious bursal disease virus (IBDV) strains is crucial for effective vaccination programs and epidemiological investigations. In this study, a combination of real-time RT-PCR and high resolution melt (HRM) curve analysis was developed for simultaneous detection and differentiation of IBDV strains/isolates. The hypervariable region of VP2 gene was amplified from several IBDV strains and subjected to HRM curve analysis. The method could readily differentiate between classical vaccines/isolates and variants. Analysis of the nucleotide sequence of the amplicons from each strain revealed that each melt curve profile was related to a unique DNA sequence. The real-time RT-PCR HRM curve analysis was also able to differentiate IBDV strains/isolates directly in bursal tissues from field submissions and from vaccinated commercial flocks. The differences between melting peaks generated from IBDV strains were significantly different (P<0.0001) demonstrating the high discriminatory power of this technique. The results presented in this study indicated that real-time RT-PCR followed by HRM curve analysis provides a rapid and robust technique for genotyping IBDV isolates/strains and can contribute to effective control of IBDV outbreaks.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Virologia
/
RNA Viral
/
Vírus da Doença Infecciosa da Bursa
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Temperatura de Transição
Tipo de estudo:
Evaluation_studies
Limite:
Animals
Idioma:
En
Revista:
J Virol Methods
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Austrália