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Title: FIELD-DEPLOYABLE REAL-TIME PCR DETECTION OF BLUETONGUE AND EPIZOOTIC HEMORRHAGIC DISEASE VIRAL RNA.

Author
item Wilson, William - Bill
item STALLKNECHT, DAVID - UNIV OF GEORGIA
item Mecham, James

Submitted to: Veterinaria Italiana
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/1/2003
Publication Date: 2/1/2005
Citation: Wilson, W.C., Stallknecht, D.E., Mecham, J.O., 2005. Field-deployable real-time pcr detection of bluetongue and epizootic hemorrhagic disease viral rna.. Veterinaria Italiana. 40(4), 587-593.

Interpretive Summary: Nucleic acid sequence information from molecular evolution studies of bluetongue virus (BTV) and related epizootic hemorrhagic disease virus (EHDV) strains has resulted in a large database of genomic information. Published sequence data and sequence data from our laboratory were used to design genetic assays for the detection of BTV or EHDV genomes. The assays can be completed in < 3 hours. The reaction conditions have been adapted to run on a field-deployable Ruggedized Advanced Pathogen Identification Device (R.A.P.I.D.) instrument from Idaho Technologies, Inc. This instrument consists of a 50-pound backpack containing everything needed to run the assays. The current assays are specific for U.S. strains of BTV and EHDV; however, new designs based on new sequencing information are being evaluated to improve specificity and sensitivity for additional serotypes. This new technology greatly enhances the speed of virus detection and the ability to monitor disease outbreaks.

Technical Abstract: Nucleic acid sequence information from molecular evolution studies of bluetongue virus (BTV) and related epizootic hemorrhagic disease virus (EHDV) strains has resulted in a large database of genomic information. Published sequence data and sequence data from our laboratory were used to design real-time field deployable RT-PCR assays for the detection of BTV or EHDV viral RNA. The assays used standard RNA extraction and TaqMan chemistries and the entire process was completed in < 3 hours. The reaction conditions have been adapted to run on a field-deployable Ruggedized Advanced Pathogen Identification Device (R.A.P.I.D.) instrument from Idaho Technologies, Inc. This instrument consists of a 50-pound backpack containing everything needed to run the assays. The current assays are specific for U.S. serotypes of BTV and EHDV; however, new designs based on new sequencing information are being evaluated to improve specificity and sensitivity for additional serotypes. This new technology greatly enhances the speed of virus detection and the ability to monitor disease outbreaks.