Next-generation whole genome sequencing of dengue virus

Methods Mol Biol. 2014:1138:175-95. doi: 10.1007/978-1-4939-0348-1_12.

Abstract

RNA viruses are notorious for their ability to quickly adapt to selective pressure from the host immune system and/or antivirals. This adaptability is likely due to the error-prone characteristics of their RNA-dependent, RNA polymerase [1, 2]. Dengue virus, a member of the Flaviviridae family of positive-strand RNA viruses, is also known to share these error-prone characteristics [3]. Utilizing high-throughput, massively parallel sequencing methodologies, or next-generation sequencing (NGS), we can now accurately quantify these populations of viruses and track the changes to these populations over the course of a single infection. The aim of this chapter is twofold: to describe the methodologies required for sample preparation prior to sequencing and to describe the bioinformatics analyses required for the resulting data.

MeSH terms

  • Base Pairing / genetics
  • Computational Biology
  • DNA, Complementary / biosynthesis
  • DNA, Viral / genetics
  • DNA, Viral / isolation & purification
  • Dengue Virus / genetics*
  • Genome, Viral / genetics*
  • High-Throughput Nucleotide Sequencing / methods*
  • Polymerase Chain Reaction
  • RNA / metabolism
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Software

Substances

  • DNA, Complementary
  • DNA, Viral
  • RNA primers
  • RNA, Viral
  • RNA