HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) An erratum has been published Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Wilson, W. C. Articles by Mecham, J. O. Search for Related Content PubMed PubMed Citation Articles by Wilson, W. C. Articles by Mecham, J. O.

Detection of all eight serotypes of Epizootic hemorrhagic disease virus by real-time reverse transcription polymerase chain reaction

Correspondence: ¹Corresponding Author: William C. Wilson, USDA, ARS, Arthropod-Borne Animal Diseases Research Laboratory, Department 3354, 1000 East University Avenue, Laramie, WY 82071, e-mail: william.wilson{at}ars.usda.gov

Epizootic hemorrhagic disease virus (EHDV) has been associated with bluetongue-like disease in cattle. Although U.S. EHDV strains have not been experimentally proven to cause disease in cattle, there is serologic evidence of infection. Differentiation of Bluetongue virus (BTV) and EHDV is necessary because diagnosis of infection caused by these viruses is often confused. The previously developed nested reverse transcription polymerase chain reaction (nRT-PCR) test for indigenous EHDV disease is sensitive and specific, but it is prone to contamination problems. Additionally, the EHDV nRT-PCR only detects 7 of the 8 serotypes. To develop an improved diagnostic test, sequence analysis was performed on 2 conserved target genes; one is highly expressed in infected mammalian cells, whereas the other is highly expressed in infected insect cells. This information was used to develop a rapid EHDV real-time PCR that detects all 8 EHDV serotypes. The EHDV assay did not cross-react with BTV strains and performed similarly to the nRT-PCR tests with archived clinical samples. In addition, it is superior to the nRT-PCR, not only because it is a closed system with fewer cross-contamination problems, but also because it detects all 8 serotypes and is less labor and time intensive.

Key Words: Bluetongue virus • Epizootic hemorrhagic disease virus • real-time reverse transcription polymerase chain reaction