title: Contamination in bluetongue virus challenge experiments.
abstract: Five cattle and five sheep that had never been exposed to bluetongue virus (BTV), as well as ten animals that had been experimentally infected with BTV-8, were inoculated with BTV-1. Previous exposure to BTV-8 did not prevent a second infection with another serotype. After the experiment, the BTV-1 preparation was found to be contaminated with BTV-15. The inoculum and blood samples taken during the experiment were analysed by serotype-specific real-time RT-PCR. There was 100-fold less BTV-15 than BTV-1 in the inoculum. Unexpectedly, BTV-15 dominated the infection in cattle that had previously been exposed to BTV-8. In sheep of both groups, on the other hand, BTV-1 prevailed over the contaminant. Regardless of the outcome, the incident demonstrates the need for a thorough contamination screening of virus preparations. For this purpose, two type-specific RT-PCR primer sets for each of the 24 established BTV serotypes as well as Toggenburg Orbivirus were designed.
authors.affiliation: Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald-Insel Riems, Germany. michael.eschbaumer@fli.bund.de
authors.forename: Michael
authors.initials: M
authors.lastname: Eschbaumer
authors.forename: Regula
authors.initials: R
authors.lastname: Wäckerlin
authors.forename: Giovanni
authors.initials: G
authors.lastname: Savini
authors.forename: Stéphan
authors.initials: S
authors.lastname: Zientara
authors.forename: Corinne
authors.initials: C
authors.lastname: Sailleau
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Bréard
authors.forename: Martin
authors.initials: M
authors.lastname: Beer
authors.forename: Bernd
authors.initials: B
authors.lastname: Hoffmann
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D001819:Bluetongue / Q000473:pathology / Q000517:prevention & control / Q000821:virology*; D001820:Bluetongue virus / Q000145:classification* / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics; D002417:Cattle; D002418:Cattle Diseases / Q000473:pathology / Q000517:prevention & control / Q000821:virology*; D002522:Chlorocebus aethiops; D017931:DNA Primers; D004866:Equipment Contamination; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012703:Serotyping; D012756:Sheep; D012757:Sheep Diseases / Q000473:pathology / Q000517:prevention & control / Q000821:virology*; D013045:Species Specificity; D014709:Vero Cells; D014765:Viral Vaccines / Q000008:administration & dosage
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D017931:DNA Primers; C442520:VP2 protein, Bluetongue virus; D014765:Viral Vaccines