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Detection of Mutation by Allele-Specific Loop-Mediated Isothermal Amplification (AS-LAMP)

  • Hiroka Aonuma
  • Athanase Badolo
  • Kiyoshi Okado
  • Hirotaka Kanuka
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1039)

Abstract

For effective control of pathogen-transmitting mosquitoes, precise surveillance data of mosquito distribution are essential. Recently, an increase of insecticide resistance due to the kdr mutation in Anopheles gambiae, a mosquito that transmits the malaria parasite, has been reported. With the aim of developing a simple and effective method for surveying resistant mosquitoes, LAMP was applied to the allele-specific detection of the kdr gene in An. gambiae. Allele-specific LAMP (AS-LAMP) method successfully distinguished the kdr homozygote from the heterozygote and the wild type. The robustness of AS-LAMP suggests its usefulness for routine identification of insects, not only mosquitoes but also other vectors and agricultural pests. Here we describe the method of AS-LAMP to detect mutation in Anopheles mosquitoes.

Key words

LAMP Mosquito Allele-specific kdr 

Notes

Acknowledgments

This study was supported in part by Grants-in-Aid for Scientific Research from the Japanese Ministry of Education, Science, Sports, Culture and Technology to H.K., and the Funding Program for the Next Generation of World-Leading Researchers (NEXT Program) to H.K. H.A. is a research fellow of the Japan Society for the Promotion of Science.

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Hiroka Aonuma
    • 1
  • Athanase Badolo
    • 2
    • 3
  • Kiyoshi Okado
    • 1
  • Hirotaka Kanuka
    • 1
  1. 1.Department of Tropical MedicineThe Jikei University School of MedicineTokyoJapan
  2. 2.University of OuagadougouOuagadougouBurkina Faso
  3. 3.Centre National de Recherche et de Formation sur le PaludismeOuagadougouBurkina Faso

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