Field-deployable real-time polymerase chain reaction detection of bluetongue and epizootic haemorrhagic disease viral ribonucleic acid.
Nucleic acid sequence information from molecular evolution studies of bluetongue virus (BTV) and related epizootic haemorrhagic 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 reverse transcriptase-polymerase chain reaction 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 <or=3 h. The reaction conditions have been adapted to run on a field-deployable ruggedised advanced pathogen identification device (RAPID) instrument from Idaho Technologies, Inc. This instrument consists of a 50-lb (22.68 kg) backpack containing everything needed to run the assays. The current assays are specific for United States 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.
Wilson
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Arthropod-Borne Animal Diseases Research Laboratory, United States Department of Agriculture, Agricultural Research Service, Laramie, WY 82071-3965, USA.
Stallknecht
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Mecham
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