Detection of mutation by allele-specific loop-mediated isothermal amplification (AS-LAMP)

Methods Mol Biol. 2013:1039:121-7. doi: 10.1007/978-1-62703-535-4_10.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Animals
  • Anopheles / genetics*
  • Base Sequence
  • DNA Primers / genetics
  • Drug Resistance / genetics
  • Genes, Insect
  • Hemizygote
  • Insecticides / pharmacology
  • Molecular Typing / methods*
  • Nephelometry and Turbidimetry
  • Nucleic Acid Amplification Techniques*

Substances

  • DNA Primers
  • Insecticides