title: IMP PCR primers detect single nucleotide polymorphisms for Anopheles gambiae species identification, Mopti and Savanna rDNA types, and resistance to dieldrin in Anopheles arabiensis.
abstract: BACKGROUND
  Polymerase chain reactions to distinguish single-nucleotide polymorphisms are commonly used for mosquito identification and identifying insecticide resistance alleles. However, the existing methods used for primer design often result in analyses that are not robust or require additional steps.
  
  
  METHODS
  Utilizing oligonucleotides that are unique in having an intentional mismatch to both templates three bases from the SNP at the 3-prime end, three new PCR assays that distinguish SNP targets using standard gel electrophoresis of undigested DNA fragments were developed and tested. These were applied to: (1) an alternative ribosomal DNA PCR assay to distinguish five members of the Anopheles gambiae complex; (2) detection of the Mopti and Savanna rDNA types; and (3) an assay to distinguish resistance to dieldrin (Rdl) alleles in Anopheles arabiensis.
  
  
  RESULTS
  Reproducible specific amplification of the target alleles was observed in all three assays. The results were consistent with existing analyses but proved simpler and the results more distinct in our hands.
  
  
  CONCLUSIONS
  The simplicity and effectiveness of the method should be utilized in these and other PCR analyses to increase their specificity and simplicity. These results have the potential to be extended not only to mosquito analyses but also to parasite and human polymorphisms.
authors.affiliation: Atlanta Research & Education Foundation, Atlanta, GA, USA. EWilkins@cdc.gov <EWilkins@cdc.gov>
authors.forename: Elien E
authors.initials: EE
authors.lastname: Wilkins
authors.forename: Paul I
authors.initials: PI
authors.lastname: Howell
authors.forename: Mark Q
authors.initials: MQ
authors.lastname: Benedict
mesh_terms: D000818:Animals; D000852:Anopheles / Q000145:classification / Q000187:drug effects / Q000235:genetics*; D017931:DNA Primers / Q000096:biosynthesis / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics*; D004026:Dieldrin / Q000494:pharmacology*; D007305:Insecticide Resistance / Q000235:genetics; D007306:Insecticides / Q000494:pharmacology*; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004275:DNA, Ribosomal; D007306:Insecticides; D004026:Dieldrin