An improved real-time polymerase chain reaction for the simultaneous detection of all serotypes of Epizootic hemorrhagic disease virus

Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
Alfonso ClavijoWilliam C Wilson

Abstract

Epizootic hemorrhagic disease virus (EHDV) is a significant pathogen of wild and sometimes domestic ungulates worldwide. Rapid and reliable methods for virus detection and identification play an essential part in the control of epizootic hemorrhagic disease (EHD). In the present study, a 1-step real-time polymerase chain reaction (PCR) group-specific assay was developed. The assay detects genome segment 5 (NS1) from all of the 8 serotypes of EHDV. Assay sensitivity was evaluated relative to a conventional gel-based nested PCR using cell culture-derived virus and diagnostic samples from clinically affected white-tailed deer (Odocoileus virginianus). The assay reliably amplified the NS1 gene from any of the EHDV strains tested, including isolates from each of the 8 EHDV serotypes. No cross-reactions were detected when all 24 serotypes of Bluetongue virus, a closely related member of the genus Orbivirus, were tested. A panel of 76 known EHDV-positive clinical samples was used to compare the performance of the assay relative to a previously reported real-time PCR assay. Results indicated that there was no statistically significant difference between the threshold cycle values obtained with both assays. A collection of 178 diagnosti...Continue Reading

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Citations

Feb 19, 2016·Transboundary and Emerging Diseases·N S MaanP P C Mertens
Oct 19, 2011·Veterinary Microbiology·Corinne SailleauStéphan Zientara
Jun 15, 2011·Research in Veterinary Science·G SaviniM Domingo
May 9, 2014·Journal of Wildlife Diseases·Feng SunAlfonso Clavijo
Oct 5, 2013·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Megan E SchroederAlfonso Clavijo
Jun 29, 2017·Journal of Wildlife Diseases·Mark G RuderDavid E Stallknecht
Apr 25, 2012·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Piet A van RijnRené G P van Gennip
Dec 29, 2020·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Kelsey T YoungJames B Stanton

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Datasets Mentioned

BETA
EU928904

Methods Mentioned

BETA
PCR
reverse transcription PCR

Software Mentioned

Primer Express

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