title: Development and evaluation of real-time polymerase chain reaction assays to identify mosquito (Diptera: Culicidae) bloodmeals originating from native Australian mammals.
abstract: Real-time TaqMan polymerase chain reaction (PCR) assays were developed for the identification of mosquito (Diptera: Culicidae) bloodmeals originating from three groups of native Australian mammals. Primers and probes were designed to amplify a partial fragment of the cytochrome b gene of the agile wallaby, Macropus agilis (Gould); brushtail possum, Trichosurus vulpecula (Kerr); and the consensus sequence of the four species of Australian flying fox: Pteropus alecto Temminck, Pteropus conspicillatus Gould, Pteropus poliocephalus Temminck, and Pteropus scapulatus Peters. When tested on DNA extracted from whole tissue, each assay was shown to be specific for the vertebrate host that it was designed to identify. To evaluate the TaqMan assays, 137 field-collected blood-fed mosquitoes were analyzed, from which 128 (93.4%) were identified using one of the assays. Compared with other PCR-based techniques for bloodmeal identification, the TaqMan assays are sensitive, specific, and provide a rapid result without the need for post-PCR manipulation and visualization of products.
authors.affiliation: Department of Microbiology and Parasitology, School of Molecular and Microbial Sciences, The University of Queensland, St. Lucia 4072, Australia. andrew_hurk@health.qld.gov.au
authors.forename: Andrew F
authors.initials: AF
authors.lastname: van den Hurk
authors.forename: Ina L
authors.initials: IL
authors.lastname: Smith
authors.forename: Greg A
authors.initials: GA
authors.lastname: Smith
mesh_terms: D000818:Animals; D001315:Australia; D001769:Blood; D009033:Culicidae / Q000502:physiology*; D045303:Cytochromes b / Q000235:genetics; D004247:DNA / Q000737:chemistry; D004063:Digestion; D005247:Feeding Behavior; D008322:Mammals / Q000145:classification / Q000235:genetics* / Q000469:parasitology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000592:standards; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D004247:DNA; D045303:Cytochromes b