A random grid based molecular epidemiological study on EBLV isolates from Germany.

Germany has reported one of the highest levels of EBLV cases in bats in Europe. So far, all isolates originating from Germany have been identified as EBLV-1, using monoclonal antibodies, and a preliminary epidemiological study has indicated that there is a distinct geographical distribution of EBLV-1 in Germany. To further investigate the spatial and temporal distribution of EBLV-1 variants in Germany and their impact on molecular epidemiology, isolates were selected using a random grid sampling procedure based on GIS. Agrid layer 30 km long over the entire area of Germany was applied to 120 geo-referenced isolates and one isolate of each grid cell containing EBLV isolates was randomly chosen. Once selected, the nucleoprotein (N) plus parts of the phosphoprotein (P) gene of each isolate were sequenced using direct cycle sequencing. Results of the subsequent phylogenetic analysis of the N-gene confirmed previous studies on European EBLVs, showing a high sequence homology between German EBLV-1 isolates. Almost identical sequence homologies within certain geographical regions indicate genomic stability during the transmission cycle of EBLV-1, with little geographic spread or intermixing. Interestingly, a 6 bp insertion as well as a single nucleotide insertion, detected in the N-P intergenic region, has been found in EBLV-1 isolates from Germany.