a comparison of parallel pyrosequencing and sanger clone-based sequencing and its impact on the characterization of the genetic diversity of hiv-1比较并行焦和桑格clone-based测序及其影响hiv - 1的基因多样性的特征.pdfVIP

A Comparison of Parallel Pyrosequencing and Sanger Clone-Based Sequencing and Its Impact on the Characterization of the Genetic Diversity of HIV-1 1 1,2 1 1 1 1 Binhua Liang *, Ma Luo , Joel Scott-Herridge , Christina Semeniuk , Mark Mendoza , Rupert Capina , 1 1 2,3 1,2 1,2 Brent Sheardown , Hezhao Ji , Joshua Kimani , Blake T. Ball , Gary Van Domselaar , Morag 1,2 1 2,4 1,2 Graham , Shane Tyler , Steven J. M. Jones , Francis A. Plummer 1 National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada, 2 Department of Medical Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada, 3 Center for STD/HIV Research and Training, University of Nairobi, Nairobi, Kenya, 4 Genome Sciences Centre, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada Abstract Background: Pyrosequencing technology has the potential to rapidly sequence HIV-1 viral quasispecies without requiring the traditional approach of cloning. In this study, we investigated the utility of ultra-deep pyrosequencing to characterize genetic diversity of the HIV-1 gag quasispecies and assessed the possible contribution of pyrosequencing technology in studying HIV-1 biology and evolution. Methodology/Principal Findings: HIV-1 gag gene was amplified from 96 patients using nested PCR. The PCR products were cloned and sequenced using capillary based Sanger fluorescent dideoxy termination sequencing. The