title: Limits of detection of bluetongue virus with different assay systems.
abstract: The sensitivity of different assay systems for detecting low concentrations of bluetongue virus (BTV) were compared. These assays included blind passage on baby hamster kidney (BHK-21) cells and on cattle pulmonary artery endothelial (CPAE) cells, immunoperoxidase staining of cells on multiwell slides, and cDNA/RNA hybridization of BTV infected cells. Nine serial 10-fold dilutions of a cell culture-adapted BTV serotype 11 were tested (each dilution was treated as a separate sample) in all assays. Visual inspection for cytopathic effects (CPE) during 3 passages in BHK-21 cells detected samples that contained greater than or equal to 3 plaque forming units (PFU)/ml of BTV. Evidence of CPE during 3 passages in CPAE cells detected samples that contained greater than or equal to 0.3 PFU/ml of BTV. A limit of detection (greater than or equal to 0.3 PFU/ml) was obtained faster by immunoperoxidase staining of BTV-inoculated CPAE cells on multiwell slides and incubated for 3 days. The cDNA/RNA hybridizations of CPAE and BHK-21 cells incubated for 2 or 3 days, respectively, with BTV dilution samples detected samples that contained greater than or equal to 30 PFU/ml. Of the assay systems examined, immunoperoxidase staining of CPAE cells on multiwell slides inoculated with cell culture-adapted BTV was the most sensitive and fastest assay for definitive virus identification.
authors.affiliation: US Department of Agriculture, Agricultural Research Service, University Station, Laramie, WY 82071-3965.
authors.forename: S J
authors.initials: SJ
authors.lastname: Wechsler
authors.forename: K J
authors.initials: KJ
authors.lastname: Austin
authors.forename: W C
authors.initials: WC
authors.lastname: Wilson
mesh_terms: D000818:Animals; D001820:Bluetongue virus / Q000235:genetics / Q000302:isolation & purification*; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D009693:Nucleic Acid Hybridization; D011237:Predictive Value of Tests; D012692:Serial Passage; D010948:Viral Plaque Assay
chemical_list: D015342:DNA Probes; D004279:DNA, Viral