While viral infections create an important threat for humanity, our knowledge of the viruses that infect humans is largely incomplete. So far, discovery of new viruses has been limited by the inability to propagate them in cell culture, the lack of serological cross-reactivity, or the absence of conserved genetic element to target by PCR. Thus, numerous acute and chronic diseases with unknown etiology are caused by yet unidentified viruses. Among the highest failure rates in determining the etiological cause of disease, pneumonia, encephalitis, and pericarditis are usually cited. In these cases, the search for unknown viruses is an urgent scientific task. We propose to identify new viruses, both indigenous and pathogenic, using metagenomics (shotgun Sanger sequencing and 454 pyrosequencing) of RNA and DNA from purified viral particles found in clinical samples of individuals presenting encephalitis, pneumonia, or pericarditis diseases without known etiology. Virus-centered bioinformatics methods will be developed to detect close as well as distant relatives of known viral species. Initial viral sequence similarity analysis will be followed by full viral genome reconstruction and phylogenetic analysis. The prevalence, abundance, and geographical distribution of newly identified viruses will then be determined using quantitative real-time PCR and RT-PCR on a 10-year collection of samples from extensively characterized patients.
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