TY - JOUR
T1 - A single fluorescence-based LAMP reaction for identifying multiple parasites in mosquitoes
AU - Aonuma, Hiroka
AU - Yoshimura, Aya
AU - Kobayashi, Tomomi
AU - Okado, Kiyoshi
AU - Badolo, Athanase
AU - Nelson, Bryce
AU - Kanuka, Hirotaka
AU - Fukumoto, Shinya
N1 - Funding Information:
We thank Tomoko Ishino, Masao Yuda, and Yasuo Chinzei for the malaria strain and Anopheles mosquito, Hiroshi Iseki for the primer design, and Kazutaka Yamada and Ryu-ichiro Maeda for the filaria and Aedes mosquito. We are also grateful to Yukari Furukawa, Emi Maekawa, Yuko Doi, Chisako Kashima, Hironori Bando, Kazuhiro Bandai, and Tokiyasu Teramoto for mosquito rearing, and Oriel Thekisoe for valuable discussion. This study was supported in part by grants from the Health Sciences Research Grant for Research on Emerging and Re-emerging Infectious Diseases from the Ministry of Health, Labor, and Welfare to H.K. and S.F., Grants-in-Aid for Scientific Research from the Japanese Ministry of Education, Science, Sports, Culture and Technology to H.K. and S.F., and the Program for the Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN) to H.K. B.N. is a research fellow of the Japan Society for the Promotion of Science.
Copyright:
Copyright 2010 Elsevier B.V., All rights reserved.
PY - 2010/6
Y1 - 2010/6
N2 - 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.
AB - 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.
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U2 - 10.1016/j.exppara.2009.12.023
DO - 10.1016/j.exppara.2009.12.023
M3 - Article
C2 - 20064511
AN - SCOPUS:77951893194
VL - 125
SP - 179
EP - 183
JO - Experimental Parasitology
JF - Experimental Parasitology
SN - 0014-4894
IS - 2
ER -