Abstract
The Variola virus is the causative agent of smallpox disease. Accurate identification of Variola and differential diagnosis between Variola and other vesicle forming (smallpox-like) pathogens are both technically challenging and of great importance. We compiled a list of vesicle forming pathogens that are prone to be misdiagnosed as smallpox. Some of the pathogens (members of the Orthopoxvirus genus) are also genetically highly similar to Variola.
We established an approach for genetic identification and differential diagnosis between the closely related Orthopoxvirus genus members. This approach couples multiplex PCR with DNA microarray as molecular means for specific and sensitive pathogen identification. It consists of PCR amplification of conserved gene fragments, followed by sequence-based specific identification on a DNA microarray platform utilizing the Arrayed Primer Extension (APEX) technique. Our prototype array – the ChiPox, is based on genes that harbor variable regions flanked by highly conserved Orthopoxvirus sequences; a pattern supporting APEX-based diagnostics. This novel approach was successfully applied for the differential identification of six members of the Orthopoxvirus genus. The ChiPox is a powerful and sensitive tool for discrimination between closely related Orthopoxvirus species, and is the first step in the establishment of a comprehensive assay for genetic discrimination of vesicle forming pathogens.
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Beth-Din, A. et al. (2010). Genetic Detection of Vesicle Forming Pathogens by Arrayed Primer Extension (APEX). In: Shafferman, A., Ordentlich, A., Velan, B. (eds) The Challenge of Highly Pathogenic Microorganisms. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9054-6_18
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DOI: https://doi.org/10.1007/978-90-481-9054-6_18
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