the sensitivity of massively parallel sequencing for detecting candidate infectious agents associated with human tissue大规模并行测序检测的敏感性候选人传染性病原体与人体组织有关.pdfVIP

The Sensitivity of Massively Parallel Sequencing for Detecting Candidate Infectious Agents Associated with Human Tissue 1 . ´ 1. 1 2 ´ 2 Richard A. Moore * , Rene L. Warren , J. Douglas Freeman , Julia A. Gustavsen , Caroline Chenard , 3 4 1 1 Jan M. Friedman , Curtis A. Suttle , Yongjun Zhao , Robert A. Holt 1 Genome Sciences Centre, BC Cancer Agency, Vancouver, British Columbia, Canada, 2 Department of Earth and Ocean Sciences, University of British Columbia, Vancouver, British Columbia, Canada, 3 Department of Medical Genetics, University of British Columbia, Child and Family Research Institute, Vancouver, British Columbia, Canada, 4 Departments of Earth and Ocean Sciences, Botany, and Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada Abstract Massively parallel sequencing technology now provides the opportunity to sample the transcriptome of a given tissue comprehensively. Transcripts at only a few copies per cell are readily detectable, allowing the discovery of low abundance viral and bacterial transcripts in human tissue samples. Here we describe an approach for mining large sequence data sets for the presence of microbial sequences. Further, we demonstrate the sensitivity of this approach by sequencing human RNA-seq libraries spiked with decreasing amounts of an RNA-virus. At a modest depth of sequencing, viral transcripts can be detected at frequencies less than 1 in 1,000,000. With current sequencing platforms approaching outputs of one billion reads per run, this is a highly sensitive method for detecting putative infectious agents a