A single fluorescence-based LAMP reaction for identifying multiple parasites in mosquitoes.
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.
Aonuma
Hiroka
H
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Yoshimura
Aya
A
Kobayashi
Tomomi
T
Okado
Kiyoshi
K
Badolo
Athanase
A
Nelson
Bryce
B
Kanuka
Hirotaka
H
Fukumoto
Shinya
S
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
D018509:DNA, Helminth; D016054:DNA, Protozoan