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Browsing by Subject "nucleic acid extraction"

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  • Suominen, Paula (University of HelsinkiHelsingin yliopistoHelsingfors universitet, 2007)
    The genus Orthopoxvirus (OPV) within the family Poxviridae Iincludes notorious smallpox virus, ectromelia virus and zoonotic cowpox, vaccinia and monkeypox viruses among others. The antibodies against OPVs are cross-reacting. Ectromelia and cowpox viruses are known to exist in Eurasia and cowpox virus also in Finland. Rodents are the reservoir hosts for both viruses and especially cowpox virus can be transmitted from rodents to humans through pet animals. In humans and cats cowpox virus causes vesicular and sometimes generalized disease. Rodents caught from Finland and Siberian Republic of Buryatia have frequently OPV-specific antibodies by immunofluorescence assay (0-92% prevalence depending on place and time). In Finland antibodies have also been demonstrated in horses, cats, dogs, bovines, lynxes and humans (Pelkonen PM et al. EID 2003). In humans the antibodies are mainly caused by smallpox vaccinations. The aims of this study were to 1) search for orthopoxvirus DNA in Finnish and Buryatian rodent panels, where OPV antibodies had been found, 2) analyze the amplified sequences to find out, which OPV species are infecting rodents and 3) deduce possible zoonotic risk. Nucleic acids were extracted from 160 Finnish and 156 Buryatian homogenized rodent lung samples by either an extraction automaton or a commercial kit. The samples were then studied with real-time PCR amplifying 150 bp piece of the hemagglutinin gene of orthopoxviruses. The rodents were selected from areas where the prevalence of OPV-specific antibodies varied from 0,9 to 69,0 %. OPV-DNA was detected from one male bank vole (Myodes glareolus) caught from Valkeakoski, Finland. The vole didn’t have detectable level of antibodies against OPV’s. The amplicon was sequenced and was shown to be identical to the hemagglutinin sequence of cowpox virus previously found in a human from Finland (Pelkonen PM et al. EID 2003). This was the first time in Finland when an ongoing OPV infection was detected in a rodent and it confirms the existence of OPVs in Finnish nature. However, some infections may have been missed due to loss in sample processing, so the true number of infected rodents might be bigger. According to the results of this study there is an existing but small risk of zoonotic orthopoxvirus infections from wild rodents in Finland and Buryatia.