Real-time RT-PCR differentiation and quantitation of infectious bursal disease virus strains using dual-labeled fluorescent probes

Real-time RT-PCR differentiation and quantitation of infectious bursal disease virus strains... A real-time RT-PCR assay was developed utilizing dual-labeled fluorescent probes binding to VP4 sequence that are specific to the classical (Cl), variant (V) and very virulent (vv) strains of infectious bursal disease virus (IBDV). The assay was highly sensitive and could detect as little as 3 × 10 2 to 3 × 10 3 copies of viral template. Viral genomic copy number could be accurately assayed over a broad range of 7–8 logs of viral genome. The variant sequence-specific probe was found to be highly specific in detecting isolates classified as variant A, D, E, G and GLS-5, and did not react with classical strains. A total of 130 field and experimental variant strain isolates were tested using this assay. The classical sequence-specific probe also demonstrated high sensitivity and specificity, and positively detected a total of 87 STC isolates, both field and experimental isolates, while differentiating between isolates that were variant and classical strains. The very virulent sequence-specific probe detected positively the Holland vvIBDV isolate and did not react with classical or variant strains. Rapid identification of viral strain is a primary concern to poultry flock health programs to ensure administered vaccines will protect against current strains of virus circulating in the flock. The ability to quantify virus concurrently is also of assistance in identifying the progression of disease outbreaks within the flock. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Virological Methods Elsevier

Real-time RT-PCR differentiation and quantitation of infectious bursal disease virus strains using dual-labeled fluorescent probes

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Publisher
Elsevier
Copyright
Copyright © 2005 Elsevier B.V.
ISSN
0166-0934
eISSN
1879-0984
D.O.I.
10.1016/j.jviromet.2005.03.009
Publisher site
See Article on Publisher Site

Abstract

A real-time RT-PCR assay was developed utilizing dual-labeled fluorescent probes binding to VP4 sequence that are specific to the classical (Cl), variant (V) and very virulent (vv) strains of infectious bursal disease virus (IBDV). The assay was highly sensitive and could detect as little as 3 × 10 2 to 3 × 10 3 copies of viral template. Viral genomic copy number could be accurately assayed over a broad range of 7–8 logs of viral genome. The variant sequence-specific probe was found to be highly specific in detecting isolates classified as variant A, D, E, G and GLS-5, and did not react with classical strains. A total of 130 field and experimental variant strain isolates were tested using this assay. The classical sequence-specific probe also demonstrated high sensitivity and specificity, and positively detected a total of 87 STC isolates, both field and experimental isolates, while differentiating between isolates that were variant and classical strains. The very virulent sequence-specific probe detected positively the Holland vvIBDV isolate and did not react with classical or variant strains. Rapid identification of viral strain is a primary concern to poultry flock health programs to ensure administered vaccines will protect against current strains of virus circulating in the flock. The ability to quantify virus concurrently is also of assistance in identifying the progression of disease outbreaks within the flock.

Journal

Journal of Virological MethodsElsevier

Published: Jul 1, 2005

References

  • A non-canonical lon proteinase lacking the ATPase domain employs the ser-Lys catalytic dyad to exercise broad control over the life cycle of a double-stranded RNA virus
    Birghan, C.; Mundt, E.; Gorbalenya, A.E.
  • The genome segment B encoding the RNA-dependent RNA polymerase protein VP1 of very virulent infectious bursal disease virus (IBDV) is phylogenetically distinct from that of all other IBDV strains
    Islam, M.R.; Zierenberg, K.; Muller, H.
  • Measuring infectious bursal disease virus RNA in blood by multiplex real-time quantitative RT-PCR
    Moody, A.; Sellers, S.; Bumstead, N.
  • Detection of infectious bursal disease virus by reverse transcription-polymerase chain reaction amplification of the virus segment A gene
    Tham, K.M.; Young, L.W.; Moon, C.D.

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