Genetic analysis of Puumala hantavirus strains from Austria

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Abstract
Sequences of the Small (S) and the Medium (M) genome segments of Puumala hantavirus (PUUV) were recovered from bank voles Clethrionomys glareolus trapped at 2 locations, Klippitztörl (Carinthia) and Ernstbrunn (Lower Austria). Lung tissue samples from 12 rodents earlier found hantavirus antibody-positive were further screened for the presence of hantaviral N-antigen using immunoblotting. RNA purified from 7 N-Ag-positive samples was subjected to the reverse transcription-polymerase chain reaction with primers designed to recover the complete S segment sequence of PUUV. The amplicons of expected size (approximately 1800 bp) have been recovered from 2 samples, 1 from Klippitztörl and another from Ernstbrunn. From the same 2 samples, PCR amplicons corresponding to the partial M segment sequence (nt 2140 to 2594), have been prepared. The S and M amplicons were sequenced and subjected to genetic analyses. Recovered hantavirus genome sequences undoubtedly belonged to PUUV genotype and the corresponding wild-type hantavirus strains were designated as PUU/Klippitztörl/Cg9/1995 and PUU/Ernstbrunn/Cg641/1995. Phylogenetic analysis placed 2 Austrian PUUV strains within a distinct genetic lineage that also included strains from the neighbouring Slovenia and Croatia. This Alpe-Adrian lineage, previously designated as 'Balkan lineage', was well supported on both S and M phylogenetic trees.