Development of polymerase chain reaction for specific identification of epizootic hemorrhagic disease virus serotype 1

I. E. Aradaib, J. W. McBride, W. C. Wilson, Bennie Osburn

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The diagnostic potential of the polymerase chain reaction (PCR) for specific identification of epizootic hemorrhagic disease virus serotype 1 (EHDV-1) in cell culture and clinical specimens was evaluated. Using oligonucleotide primers, selected from genome segment 2 of EHDV-1 (New Jersey strain), the PCR-based assay resulted in a 862 base pair (bp) PCR product. EHDV-1 RNA from United States prototype serotype 1 and a number of EHDV-1 field isolates, propagated in cell cultures, were detected by this PCR based assay. The specific 862 bp PCR products were visualized on ethidium bromide-stained agarose gel. Identity of the PCR product was confirmed by chemiluminescent hybridization with non radiolabelled internal probe. Using chemiluminescent hybridization, the sensitivity of the PCR assay was 1.0 fg of virus RNA (equivalent to 60 virus particles). Amplification product was not detected when the PCR-based assay was applied to RNA from EHDV serotype 2 (EHDV-2); the United States bluetongue virus (BLU) prototypes serotypes 2, 10, 11, 13, and 17; total nucleic acid extracts from uninfected BHK-21 cell; or blood cells from calves and deer that were EHDV-seronegative and virus isolation negative. Application of this EHDV-1 PCR-based assay to clinical samples resulted in detection of EHDV-1 RNA from blood samples, collected from a calf experimentally infected with EHDV-1. The described PCR-based assay provides a simple, rapid, sensitive, specific and inexpensive method for specific identification of EHDV-1 infection in susceptible ruminants.

Original languageEnglish (US)
Pages (from-to)2273-2281
Number of pages9
JournalArchives of Virology
Volume140
Issue number12
DOIs
StatePublished - Dec 1 1995

Fingerprint

Epizootic hemorrhagic disease virus
Polymerase Chain Reaction
RNA
Base Pairing
Serogroup
Cell Culture Techniques
Bluetongue virus
DNA Primers
Deer
Ethidium
RNA Viruses
Ruminants
Virion
Sepharose
Nucleic Acids

ASJC Scopus subject areas

  • Virology

Cite this

Development of polymerase chain reaction for specific identification of epizootic hemorrhagic disease virus serotype 1. / Aradaib, I. E.; McBride, J. W.; Wilson, W. C.; Osburn, Bennie.

In: Archives of Virology, Vol. 140, No. 12, 01.12.1995, p. 2273-2281.

Research output: Contribution to journalArticle

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