TY - JOUR T1 - Long single-molecule reads can resolve the complexity of the Influenza virus composed of rare, closely related mutant variants JF - bioRxiv DO - 10.1101/036392 SP - 036392 AU - Alexander Artyomenko AU - Nicholas C Wu AU - Serghei Mangul AU - Eleazar Eskin AU - Ren Sun AU - Alex Zelikovsky Y1 - 2016/01/01 UR - http://biorxiv.org/content/early/2016/04/28/036392.abstract N2 - As a result of a high rate of mutations and recombination events, an RNA-virus exists as a heterogeneous “swarm” of mutant variants. The long read length offered by single-molecule sequencing technologies allows each mutant variant to be sequenced in a single pass. However, high error rate limits the ability to reconstruct heterogeneous viral population composed of rare, related mutant variants. In this paper, we present 2SNV, a method able to tolerate the high error-rate of the single-molecule protocol and reconstruct mutant variants. 2SNV uses linkage between single nucleotide variations to efficiently distinguish them from read errors. To benchmark the sensitivity of 2SNV, we performed a single-molecule sequencing experiment on a sample containing a titrated level of known viral mutant variants. Our method is able to accurately reconstruct clone with frequency of 0.2% and distinguish clones that differed in only two nucleotides distantly located on the genome. 2SNV outperforms existing methods for full-length viral mutant reconstruction. The open source implementation of 2SNV is freely available for download at http://alan.cs.gsu.edu/NGS/?q=content/2snv ER -