A simple restriction fragment PCR approach for discrimination of humanpathogenic Old World animal Orthopoxvirus species

Authors: Huemer, Hartwig P.; Hönlinger, Bettina; Höpfl, Reinhard

Source: Canadian Journal of Microbiology, Volume 54, Number 2, February 2008 , pp. 159-162(4)

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Abstract:

There are reliable polymerase chain reaction assays available for exclusion of Variola virus from other poxviruses. However, the discrimination of humanpathogenic animal Orthopoxviridae is more challenging because of the high genomic conservation. Based on the variability of the A36R gene, we describe a simple 20 min PCR assay followed by a 1 h digest with 3 different restriction enzymes. This assay enables rapid discrimination between Cowpox virus and Monkeypox virus and discrimination of the most prevalent members of the Vaccinia virus and Camelpox virus. The test was orthopoxvirus specificand did not react with parapox (Orf) virus. Moreover, the amplified fragments were also well suited for additional genotyping by direct DNA sequencing.

Il existe des essais en PCR fiables qui permettent d'identifier le virus de la variole (Variola) parmi les autres poxvirus. Cependant, il est plus difficile de discriminer les Orthopoxviridae pathogènes pour l'humain et l'animal à cause d'un haut niveau de conservation génomique. Nous décrivons un essai PCR simple de 20 min, basé sur la variabilité du gène A36R, suivi d'une digestion d'une heure avec trois enzymes de restriction différentes. Ceci permet une discrimination rapide des virus de la variole bovine et simienne (Cowpox virus et Monkeypox virus), ainsi que celle des virus dont la prévalence est la plus élevée, soit les virus de la vaccine et de la variole des camélidés (Camelpox virus). Le test est spécifique aux virus orthopox, ne réagissant pas aux virus parapox (Orf). De plus, les fragments amplifiés conviennent aussi au génotypage par séquençage direct d'ADN.

Document Type: Research article

Publication date: 2008-02-02

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Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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A simple restriction fragment PCR approach for discrimination of humanpathogenic Old World animal Orthopoxvirus species

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