title: Development of an allele-specific, loop-mediated, isothermal amplification method (AS-LAMP) to detect the L1014F kdr-w mutation in Anopheles gambiae s. l.
abstract: BACKGROUND
  Malaria 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.
  
  
  METHODS
  DNA 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.
  
  
  RESULTS
  The 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.
  
  
  CONCLUSIONS
  The AS-LAMP method can be used to detect the kdr mutation for quick decision-making, even in less well-equipped laboratories.
authors: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido, 080-8555, Japan. a.badolo@gmail.com
authors: Athanase
authors: A
authors: Badolo
authors: Kyioshi
authors: K
authors: Okado
authors: Wamdaogo M
authors: WM
authors: Guelbeogo
authors: Hiroka
authors: H
authors: Aonuma
authors: Hironori
authors: H
authors: Bando
authors: Shinya
authors: S
authors: Fukumoto
authors: N'Fale
authors: N
authors: Sagnon
authors: Hirotaka
authors: H
authors: Kanuka
mesh_terms: D000483:Alleles; D000818:Animals; D000852:Anopheles / Q000235:genetics*; D017931:DNA Primers; D004199:Disease Vectors; D004772:Entomology / Q000379:methods*; D007305:Insecticide Resistance; D020125:Mutation, Missense; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D015222:Sodium Channels / Q000235:genetics*; D013696:Temperature
chemical_list: D017931:DNA Primers; D015222:Sodium Channels