Development of an allele-specific, loop-mediated, isothermal amplification method (AS-LAMP) to detect the L1014F kdr-w mutation in Anopheles gambiae s. l.

Malaria Journal

Methodology

N’Fale Sagnon¹  Wamdaogo M Guelbeogo¹  Shinya Fukumoto²  Hiroka Aonuma³  Kyioshi Okado³  Hirotaka Kanuka³  Hironori Bando³  Athanase Badolo⁴ 

[1] Centre National de Recherche et de Formation sur le Paludisme, BP 2208, Ouagadougou 01, Burkina Faso;National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, 080-8555, Inada-cho, Obihiro, Hokkaido, Japan;National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, 080-8555, Inada-cho, Obihiro, Hokkaido, Japan;Department of Tropical Medicine, The Jikei University School of Medicine, 3-25-8, Nishi-Shinbashi, 105-8461, Minato-ku, Tokyo, Japan;National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, 080-8555, Inada-cho, Obihiro, Hokkaido, Japan;University of Ouagadougou, BP 7021, Ouagadougou 03, Burkina Faso;Centre National de Recherche et de Formation sur le Paludisme, BP 2208, Ouagadougou 01, Burkina Faso;

关键词: Insecticide;    Resistance;    kdr;    LAMP;    Malaria;    Mosquito;   

DOI  :  10.1186/1475-2875-11-227

 received in 2012-04-06, accepted in 2012-07-06,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundMalaria control relies heavily on treated bed nets and indoor residual spraying with pyrethroid insecticides. Unfortunately, the resistance to pyrethroid insecticides, mainly due to the kdr mutation, is spreading in the main malaria vector Anopheles gambiae s.l., decreasing the insecticides’ efficacy. To manage the insecticide resistance rapidly and flexibly, simple and effective tools for the early detection of resistant mosquitoes are needed. This study aimed to develop an allele-specific, loop-mediated, isothermal amplification (AS-LAMP) method to detect the West African-type kdr mutation (kdr-w; L1014F) in field-collected mosquitoes.MethodsDNA fragments of the wild-type and the mutated kdr gene were used to select the primers and develop the method. The primers were designed with the mutation at the 5’ end of the backward inner primer (BIP). The AS-LAMP method was compared to the AS-PCR method using the genomic DNA of 120 field-collected mosquitoes.ResultsThe AS-LAMP method could discriminate between the wild-type homozygote, the heterozygote, and the kdr-w homozygote within 75 min. The AS-LAMP method has the advantage of being faster and at least as sensitive and specific as the AS-PCR method.ConclusionsThe AS-LAMP method can be used to detect the kdr mutation for quick decision-making, even in less well-equipped laboratories.

【 授权许可】

CC BY   
© Badolo et al.; licensee BioMed Central Ltd. 2012