A multiplex PCR to differentiate the two sibling species of mosquitoes Ochlerotatus detritus and Oc. coluzzii and evidence for further genetic heterogeneity within the Detritus complex

Infect Genet Evol. 2014 Dec:28:676-80. doi: 10.1016/j.meegid.2014.07.010. Epub 2014 Jul 18.

Abstract

Internal transcribed spacer regions of ribosomal DNA were sequenced, and species-specific primers were designed to simplify the identification of two morphologically similar species of the Detritus complex, Ochlerotatus detritus and Ochlerotatuscoluzzii. Each newly designed primer was able to amplify a species-specific fragment with a different size. Samples from France and Germany were successfully tested. This new tool prompts for bio-ecological studies to refine basic knowledge on the bionomics of this species complex, towards a better control and prevention of ensuing nuisances. Moreover, ITS2 sequencing revealed the existence of (1) two distinct haplotypes of Oc. detritus that are sympatric and widely distributed along the French Atlantic and Mediterranean littorals and (2) a specific haplotype in mosquitoes sampled from Tunisia, raising the question of the taxonomic status of this North-African population.

Keywords: Aedini; Mosquito; Multiplex PCR; Species determination.

MeSH terms

  • Alleles
  • Animals
  • Base Sequence
  • DNA, Mitochondrial / chemistry
  • DNA, Mitochondrial / genetics
  • DNA, Ribosomal Spacer / chemistry
  • DNA, Ribosomal Spacer / genetics
  • France
  • Genetic Heterogeneity*
  • Germany
  • Haplotypes
  • Male
  • Molecular Sequence Data
  • Multiplex Polymerase Chain Reaction
  • Ochlerotatus / classification*
  • Ochlerotatus / genetics*
  • Phylogeny
  • Reproducibility of Results
  • Sequence Alignment

Substances

  • DNA, Mitochondrial
  • DNA, Ribosomal Spacer