Species-level identification of orthopoxviruses with an oligonucleotide microchip - PubMed (original) (raw)

doi: 10.1128/JCM.40.3.753-757.2002.

Maxim Mikheev, Sergei Shchelkunov, Vladimir Mikhailovich, Alexander Sobolev, Vladimir Blinov, Igor Babkin, Alexander Guskov, Elena Sokunova, Alexander Zasedatelev, Lev Sandakhchiev, Andrei Mirzabekov

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Species-level identification of orthopoxviruses with an oligonucleotide microchip

Sergey Lapa et al. J Clin Microbiol. 2002 Mar.

Abstract

A method for species-specific detection of orthopoxviruses pathogenic for humans and animals is described. The method is based on hybridization of a fluorescently labeled amplified DNA specimen with the oligonucleotide DNA probes immobilized on a microchip (MAGIChip). The probes identify species-specific sites within the crmB gene encoding the viral analogue of tumor necrosis factor receptor, one of the most important determinants of pathogenicity in this genus of viruses. The diagnostic procedure takes 6 h and does not require any sophisticated equipment (a portable fluorescence reader can be used).

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Figures

FIG. 1.

FIG. 1.

Region of the variola virus vTNFR gene used as a DNA target for on-chip hybridization analysis. The sequences of oligonucleotide probes 1 to 15 and their locations relative to the crmB gene sequence are shown. The species-specific nucleotides are underlined. The dashes indicate deletions.

FIG. 2.

FIG. 2.

Hybridization patterns of six OPV DNAs from variola minor virus strain Garcia-1966 (a), vaccinia virus strain Wyeth (b), monkeypox virus strain Zaire-96 (c), vaccinia virus strain rabbitpox Utrecht (d), cowpox virus strain GRI-90 (e), and camelpox virus strain CP-1 (f). In the graphs on the left, the expected fluorescence patterns are shown. The numbers inside the microchip elements correspond to the probe numbering in Fig. 1.

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