title: A single fluorescence-based LAMP reaction for identifying multiple parasites in mosquitoes.
abstract: Vector-borne diseases, such as malaria and lymphatic filariasis, are co-endemic in large parts of the world. To develop a multiplex amplification method for the simultaneous detection of multiple insect-borne infectious diseases, we used LAMP with fluorescently labeled primers to identify the SPECT2 gene of Plasmodium berghei and the cytochrome oxidase subunit I gene of Dirofilaria immitis in mosquitoes. This technique could detect as few as 100 P. berghei-infected red blood cell-equivalents or one D. immitis microfilaria. Moreover, individual species of parasites in mosquitoes could be identified when a mixture of fluorescently labeled primer sets was used. These findings suggest that the multiplex LAMP assay is sensitive and specific enough to identify parasite-bearing mosquitoes in areas where several diseases occur simultaneously. This procedure could increase the efficiency and effectiveness of arthropod-borne disease elimination programs.
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
authors.forename: Hiroka
authors.initials: H
authors.lastname: Aonuma
authors.forename: Aya
authors.initials: A
authors.lastname: Yoshimura
authors.forename: Tomomi
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Okado
authors.forename: Athanase
authors.initials: A
authors.lastname: Badolo
authors.forename: Bryce
authors.initials: B
authors.lastname: Nelson
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Kanuka
authors.forename: Shinya
authors.initials: S
authors.lastname: Fukumoto
mesh_terms: D000330:Aedes / Q000469:parasitology*; D000818:Animals; D000852:Anopheles / Q000469:parasitology*; D018509:DNA, Helminth / Q000302:isolation & purification; D016054:DNA, Protozoan / Q000302:isolation & purification; D004183:Dirofilaria immitis / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D007303:Insect Vectors / Q000469:parasitology*; D008317:Malpighian Tubules / Q000469:parasitology; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D021141:Nucleic Acid Amplification Techniques / Q000379:methods* / Q000592:standards; D010962:Plasmodium berghei / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
chemical_list: D018509:DNA, Helminth; D016054:DNA, Protozoan