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Sequences of wild Puumala virus genes show a correlation of genetic variation with geographic origin of the strains

1994; Microbiology Society; Volume: 75; Issue: 2 Linguagem: Inglês

10.1099/0022-1317-75-2-405

1465-2099

Alexander Plyusnin, Olli Vapalahti, K. Ulfves, Heikki Lehväslaiho, N. S. Apekina, I. N. Gavrilovskaya, V. Blinov, A. Vaheri,

Viral Infections and Outbreaks Research

An experimental scheme was developed for direct sequence analysis of Puumala virus-containing specimens from wild rodents (Clethrionomys glareolus). Total RNA isolated from rodent lung tissues was reverse-transcribed in the presence of a universal 11 nucleotide primer complementary to all three viral RNA segments followed by amplification in a PCR with gene-specific primers. A full-length PCR product of approximately 1800 bp from the S segment encoding the viral nucleo-protein and a product of approximately 900 bp from the M segment (encoding the C-terminal two-thirds of the G2 protein and including the 3′ non-coding region) of Puumala virus (from C. glareolus trapped in Udmurtia) were prepared and sequenced. No pronounced differences to Vero cell-grown viruses were seen. The Udmurtia/894Cg/91 strain was more closely related to the Bashkiria/CG 18–20/84 strain than to the Finnish prototype strain of Puumala virus, Sotkamo/V-2969/81. Thus there is a correlation with the geographic origin of the three strains. The results indicate the occurrence of genetic drift and different selection pressures leading to (i) clustering of mutations, (ii) a lower frequency of nucleotide substitutions in the coding than in the 3′ noncoding regions and (iii) a higher frequency of amino acid substitutions in G2 than in the N protein.