Phylogenetic analysis of the 5′-untranslated region of German BVDV type II isolates

On the basis of genetic differences, bovine viral diarrhoea viruses (BVDV) are subclassified into two distinct genotypes, BVDV type I and BVDV type II. We selected German BVDV type II isolates using the BVDV type I-specific monoclonal antibody WB160 and flow cytometric analysis for further characterization. For molecular characterization, a 288-by fragment of the 5'-untranslated region (5'-UTR) of the selected isolates was investigated by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing. Sequence comparisons of the partial 5'-UTR sequences and their phylogenetic analyses demonstrated that the 18 German BVDV type II isolates all belong to either subtype IIa (10 isolates) or subtype IIc (eight isolates). Nevertheless, the German BVDV type II isolates were genetically different (89.9-94.3% sequence identity) from the standard BVDV type II strain 890 from North America, which was recently classified as BVDV type IIa. Furthermore, a clear subdivision of the German BVDV type II isolates into two distinct subtypes (BVDV IIa Germany and BVDV IIc Germany) is shown. Viruses of both subgroups differed in the analysed 5'-UTR fragment from each other (91.6-95.2% sequence identity), but were highly conserved within the same German subtype (97.2-100% sequence identity). These findings are discussed in the context of BVDV type II origin, possible introduction into Germany, its epidemiology and impact for vaccine development.