Application of the Droplet Digital Polymerase Chain Reaction (ddPCR) Platform for Detection and Quantification of Vertebrate Host DNA in Engorged Mosquitoes

L M Rice; L L Robb; D A Hartman; J R Anderson; R C Kading

doi:10.1093/jme/tjz016

Application of the Droplet Digital Polymerase Chain Reaction (ddPCR) Platform for Detection and Quantification of Vertebrate Host DNA in Engorged Mosquitoes

J Med Entomol. 2019 Jun 27;56(4):1150-1153. doi: 10.1093/jme/tjz016.

Authors

L M Rice¹, L L Robb¹, D A Hartman¹, J R Anderson¹, R C Kading¹

Affiliation

¹ Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO.

PMID: 30856248
DOI: 10.1093/jme/tjz016

Abstract

Hematophagous arthropod bloodmeal identification has remained a challenge in the field of vector biology, but these studies are important to understand blood feeding patterns of arthropods, spatial, and temporal patterns in arbovirus transmission cycles, and risk of human and veterinary disease. We investigated the use of an existing vertebrate primer set for use on the droplet digital polymerase chain reaction (ddPCR) platform, to explore the use of this technology in the identification and quantification of vertebrate DNA in mosquito blood meals. Host DNA was detectable 48-h post-engorgement in some mosquitoes by ddPCR, compared with 24-h post-engorgement using traditional PCR. The capability of ddPCR for absolute quantification of template DNA offers unique potential applications of this new technology to field studies on the ecology of vector-borne diseases, but currently with limited scope.

Keywords: detection; diagnostics; feeding behavior; mosquito-borne disease; survey.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cattle
Culicidae / chemistry*
DNA / analysis*
Polymerase Chain Reaction

Substances

DNA