Barcoding blood meals: New vertebrate-specific primer sets for assigning taxonomic identities to host DNA from mosquito blood meals

PLoS Neglected Tropical Diseases
Lawrence E ReevesPhillip E Kaufman

Abstract

The transmission dynamics of mosquito-vectored pathogens are, in part, mediated by mosquito host-feeding patterns. These patterns are elucidated using blood meal analysis, a collection of serological and molecular techniques that determine the taxonomic identities of the host animals from which blood meals are derived. Modern blood meal analyses rely on polymerase chain reaction (PCR), DNA sequencing, and bioinformatic comparisons of blood meal DNA sequences to reference databases. Ideally, primers used in blood meal analysis PCRs amplify templates from a taxonomically diverse range of vertebrates, produce a short amplicon, and avoid co-amplification of non-target templates. Few primer sets that fit these requirements are available for the cytochrome c oxidase subunit I (COI) gene, the species identification marker with the highest taxonomic coverage in reference databases. Here, we present new primer sets designed to amplify fragments of the DNA barcoding region of the vertebrate COI gene, while avoiding co-amplification of mosquito templates, without multiplexed or nested PCR. Primers were validated using host vertebrate DNA templates from mosquito blood meals of known origin, representing all terrestrial vertebrate classes, ...Continue Reading

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Citations

Apr 14, 2020·Journal of Medical Entomology·Richard G WestNathan D Burkett-Cadena
Jul 2, 2020·PeerJ·Pedro M PedroMaria Anice Mureb Sallum
Jan 10, 2021·Insects·Erin M Borland, Rebekah C Kading
Jul 21, 2021·Parasite : Journal De La Société Française De Parasitologie·Zuzana KasičováAlica Kočišová
Aug 17, 2021·PeerJ·Vasco ElbrechtDirk Steinke
Dec 5, 2021·Molecular Ecology Resources·Lorenz WührlRudolf Meier
Nov 10, 2020·Acta Tropica·Sandra Sayuri NagakiMaria Anice Mureb Sallum

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Methods Mentioned

BETA
PCR
electrophoresis
PCRs

Software Mentioned

R
R package Agricolae
BOLD
Geneious
Basic Local Alignment Search Tool ( BLAST )
stats

Related Concepts

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Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.