A Microarray-Based Assay for the Detection of Viruses in Municipal Drinking Water (CBX87). unidentified

Viruses are the most abundant biological entities in natural waters. In addition to causing disease, they play a dominant role in biogeochemical cycling and serve as a major genetic reservoir for driving evolution. Human viral infections from drinking water in the United States are estimated at ~6.5 million cases per year, more than 10 times higher than those caused by bacteria or protozoans. The U.S. Environmental Protection Agency mandates the monitoring of a handful of the estimated 100 human enteric viruses and several viral indicators, but monitoring is only required when an outbreak occurs, mainly because of the lack of viral detection capabilities at most water quality laboratories across the nation. To begin to address this need, this paper describes the use of a high-throughput screening tool for simultaneous detection of thousands of viral genes within municipal water. Viruses were sampled and concentrated from several municipal drinking water sources. Total viral nucleic acids were extracted following filtration. After amplification, viral DNA and cDNA were pooled, fluorescently labeled, and hybridized onto a Combimatrix? 12K microarray containing probes for >1,000 human viruses. Nucleic acid signatures from the Reoviridae, Mononegavirales, Retroviridae, Adenoviridae, Nidovirales, Herpesviridae, Papillomaviridae, Reoviridae, Orthomyxoviridae, Tombusviridae, Arenaviridae, and Poxviridae families exhibited positive hybridizations. Quantitative PCR for genetic markers of the Adenoviridae and Orthomyxoviridae families confirmed the microarray results. This study describes the first use and validation of a microarray-based assay for the detection of viruses within municipal water supplies.