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.
| Original language | English |
|---|---|
| Pages (from-to) | 264-271 |
| Number of pages | 8 |
| Journal | Journal of Virological Methods |
| Volume | 171 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 01 Jan 2011 |
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Differentiation of infectious bursal disease virus strains using real-time RT-PCR and high resolution melt curve analysis. / Ghorashi, Seyed A.; O'Rourke, Denise; Ignjatovic, Jagoda; Noormohammadi, Amir H.
In: Journal of Virological Methods, Vol. 171, No. 1, 01.01.2011, p. 264-271.Research output: Contribution to journal › Article
TY - JOUR
T1 - Differentiation of infectious bursal disease virus strains using real-time RT-PCR and high resolution melt curve analysis
AU - Ghorashi, Seyed A.
AU - O'Rourke, Denise
AU - Ignjatovic, Jagoda
AU - Noormohammadi, Amir H.
PY - 2011/1/1
Y1 - 2011/1/1
N2 - 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.
AB - 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.
KW - Genotyping
KW - High resolution melt curve analysis
KW - Infectious bursal disease virus
KW - Strain differentiation
UR - http://www.scopus.com/inward/record.url?scp=78650534658&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=78650534658&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2010.11.013
DO - 10.1016/j.jviromet.2010.11.013
M3 - Article
C2 - 21111004
AN - SCOPUS:78650534658
VL - 171
SP - 264
EP - 271
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 1
ER -