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PubMed · 16690139

A multiplex DNA suspension microarray for simultaneous detection and differentiation of classical swine fever virus and other pestiviruses.

Dirk Deregt·Scott A Gilbert·Sandor Dudas·John Pasick·Shailja Baxi·Kimberley M Burton·Mohit K Baxi
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Abstract

An oligonucleotide suspension microarray (Luminex microsphere system) was developed for detection and differentiation of animal pestiviruses: classical swine fever virus (CSFV), bovine viral diarrhea virus types 1 and 2 (BVDV1 and BVDV2), and border disease virus (BDV). Species-specific and pestivirus-common oligonucleotide probes were designed to the 5' UTR region and conjugated to individual color-coded Luminex carboxy beads (probe beads). Target pestivirus sequences were amplified by asymmetric PCR using a biotinylated reverse primer and a forward and reverse primer ratio of 1:5. The biotinylated products were hybridized to eight probe beads in a multiplex assay and analyzed using streptavidin conjugated to a fluorescent reporter molecule. The assay was able to detect and differentiate all 40 strains of CSFV, BVDV1, BVDV2 and BDV tested. The analytical sensitivity was determined to be 0.2-10 TCID50/ml. The major advantages of the DNA-microsphere suspension microarray, as a low density array, are its ease of handling and ability to simultaneously detect and type multiple infectious agents.

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5' Untranslated RegionsBiotinFluorescenceGenotypeMicrospheresOligonucleotide Array Sequence AnalysisPestivirusPolymerase Chain ReactionRNA, ViralSensitivity and SpecificityStaining and LabelingStreptavidinJournal ArticleResearch Support, Non-U.S. Gov't

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Dirk Deregt, Scott A Gilbert, Sandor Dudas, John Pasick, Shailja Baxi, Kimberley M Burton, Mohit K Baxi. 2006-05-09. A multiplex DNA suspension microarray for simultaneous detection and differentiation of classical swine fever virus and other pestiviruses.. https://doi.org/10.1016/j.jviromet.2006.03.025

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