Simultaneous identification of host, ectoparasite and pathogen DNA via in-solution capture

Mol Ecol Resour. 2016 Sep;16(5):1224-39. doi: 10.1111/1755-0998.12524. Epub 2016 Apr 4.

Abstract

Ectoparasites frequently vector pathogens from often unknown pathogen reservoirs to both human and animal populations. Simultaneous identification of the ectoparasite species, the wildlife host that provided their most recent blood meal(s), and their pathogen load would greatly facilitate the understanding of the complex transmission dynamics of vector-borne diseases. Currently, these identifications are principally performed using multiple polymerase chain reaction (PCR) assays. We developed an assay (EctoBaits) based on in-solution capture paired with high-throughput sequencing to simultaneously identify ectoparasites, host blood meals and pathogens. We validated our in-solution capture results using double-blind PCR assays, morphology and collection data. The EctoBaits assay effectively and efficiently identifies ectoparasites, blood meals, and pathogens in a single capture experiment, allowing for high-resolution taxonomic identification while preserving the DNA sample for future analyses.

Keywords: DNA; blood meal; ectoparasite; high-throughput sequencing; in-solution capture; pathogen.

MeSH terms

  • Animals
  • DNA / genetics
  • DNA / isolation & purification
  • Ecosystem*
  • Genotyping Techniques / methods*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Molecular Biology / methods*
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA

Substances

  • DNA

Associated data

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