Characterization of a Newly Discovered Picornavirus from a Patient with Diarrhea

The overall goal of this project is to complete our characterization of a novel picornavirus that we discovered in a pediatric diarrhea sample. <P> Diarrhea is the third leading infectious cause of death worldwide and causes approximately 2 million deaths and 1.4 billion non-fatal episodes yearly. It is estimated that 40% of diarrhea cases are of unknown etiology. A conservative estimate indicates that at least 500,000 annual deaths are caused by pediatric diarrhea of unknown etiology, and therefore the identification of a novel agent responsible for even a fraction of these episodes would be of great significance to human health and fundamental science. <P> We recently conducted a viral metagenomic analysis of diarrhea samples using mass sequencing. In a stool collected in Melbourne, Australia from a child with acute diarrhea, one 395 bp sequence read was identified that had 55% amino acid identity to a known picornavirus protein, suggesting that a novel picornavirus was present in this sample, which we have termedWUPV. <P> The goal of this project is to complete the sequence of this novel picornavirus, determine the extent to which it is found in additional specimens, determine if WUPV is associated with diarrhea, and generate an infectious clone or culture the virus. The sequence of the genome will be completed by 5' random amplification of cDNA ends (RACE) from RNA extracted from the stool sample. <P> After the full sequence has been obtained an infectious clone will be generated. Additionally, we will attempt to culture the virus in a variety of different cell lines. <P> To determine if WUPV is present in other samples we will use different pairs of specific primers to screen 3 international and local collections of stools. <P> To demonstrate if WUPV is associated with diarrhea we will use these same specific primers on a cohort of diarrhea stools and control stools that are age-, geographic- and season- matched to each of the diarrhea specimens. <P> Epidemiological and biological characterization of our newly discovered picornavirus will confirm or refute its association with, and potential contribution to, the global disease burden of gastroenteritis.