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Simultaneous identification of host, ectoparasite and pathogen DNA via in‐solution capture
Author(s) -
Campana Michael G.,
Hawkins Melissa T. R.,
Henson Lauren H.,
Stewardson Kristin,
Young Hillary S.,
Card Leah R.,
Lock Justin,
Agwanda Bernard,
Brinkerhoff Jory,
Gaff Holly D.,
Helgen Kristofer M.,
Maldonado Jesús E.,
McShea William J.,
Fleischer Robert C.
Publication year - 2016
Publication title -
molecular ecology resources
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.96
H-Index - 136
eISSN - 1755-0998
pISSN - 1755-098X
DOI - 10.1111/1755-0998.12524
Subject(s) - biology , pathogen , host (biology) , polymerase chain reaction , identification (biology) , blood meal , vector (molecular biology) , computational biology , dna sequencing , dna , virology , microbiology and biotechnology , recombinant dna , genetics , zoology , gene , ecology
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.