HIV-1 quasispecies delineation by tag linkage deep sequencing.

• Main Authors: Nicholas C Wu, Justin De La Cruz, Laith Q Al-Mawsawi, C Anders Olson, Hangfei Qi, Harding H Luan, Nguyen Nguyen, Yushen Du, Shuai Le, Ting-Ting Wu, Xinmin Li, Martha J Lewis, Otto O Yang, Ren Sun
• Format: Article
• Language: English
• Published: Public Library of Science (PLoS) 2014-01-01
• Series: PLoS ONE
• Online Access: http://europepmc.org/articles/PMC4026136?pdf=render

Trade-offs between throughput, read length, and error rates in high-throughput sequencing limit certain applications such as monitoring viral quasispecies. Here, we describe a molecular-based tag linkage method that allows assemblage of short sequence reads into long DNA fragments. It enables haplotype phasing with high accuracy and sensitivity to interrogate individual viral sequences in a quasispecies. This approach is demonstrated to deduce ∼ 2000 unique 1.3 kb viral sequences from HIV-1 quasispecies in vivo and after passaging ex vivo with a detection limit of ∼ 0.005% to ∼ 0.001%. Reproducibility of the method is validated quantitatively and qualitatively by a technical replicate. This approach can improve monitoring of the genetic architecture and evolution dynamics in any quasispecies population.