A sensitive, specific and reproducible real-time polymerase chain reaction method for detection of Plasmodium vivax and Plasmodium falciparum infection in field-collected anophelines

Mem Inst Oswaldo Cruz. 2015 Jun;110(4):573-6. doi: 10.1590/0074-02760150031. Epub 2015 May 29.

Abstract

We describe a simple method for detection of Plasmodium vivax and Plasmodium falciparum infection in anophelines using a triplex TaqMan real-time polymerase chain reaction (PCR) assay (18S rRNA). We tested the assay on Anopheles darlingi and Anopheles stephensi colony mosquitoes fed with Plasmodium-infected blood meals and in duplicate on field collected An. darlingi. We compared the real-time PCR results of colony-infected and field collected An. darlingi, separately, to a conventional PCR method. We determined that a cytochrome b-PCR method was only 3.33% as sensitive and 93.38% as specific as our real-time PCR assay with field-collected samples. We demonstrate that this assay is sensitive, specific and reproducible.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Anopheles / parasitology*
  • Cytochromes b / genetics
  • Insect Vectors / parasitology*
  • Plasmodium falciparum / genetics*
  • Plasmodium falciparum / isolation & purification
  • Plasmodium vivax / genetics*
  • Plasmodium vivax / isolation & purification
  • Real-Time Polymerase Chain Reaction*
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Cytochromes b