Plasmodium vivax: parasitemia determination by real-time quantitative PCR in Aotus monkeys

Exp Parasitol. 2002 Feb;100(2):131-4. doi: 10.1016/S0014-4894(02)00010-3.


Plasmodium vivax and Plasmodium falciparum are the two prevalent human malaria species. A Colombian P. vivax wild strain has been adapted in Aotus nancymaae monkeys for use in further biological and immunological studies. We present data validating a real-time PCR assay quantifying P. vivax parasitemia, using the small subunit ribosomal RNA genes as an amplification target. P. vivax species-specific primers were designed on the 18S ribosomal gene V8 region, for amplifying both asexual and sporozoite ssrRNA genes. The assay detects amplification products bound to fluorescent SYBR-Green I dye using Perkin-Elmer GeneAmp-5700-SDS. Linear range standard curves from 6 DNA concentration logs (+0.99 correlation coefficients) were obtained. Standard curves were constructed using a plasmid containing target gene for real-time PCR amplification. This P. vivax specific assay is very sensitive, having a three parasite detection limit, and is reproducible and accurate. It involves a "closed-tube" PCR, avoids time-consuming post-PCR manipulation, and decreases potential PCR contamination.

Publication types

  • Validation Study

MeSH terms

  • Animals
  • Aotidae
  • DNA, Protozoan / blood
  • DNA, Ribosomal / blood*
  • Malaria, Vivax / diagnosis*
  • Parasitemia / diagnosis*
  • Plasmodium vivax / genetics
  • Plasmodium vivax / isolation & purification*
  • Polymerase Chain Reaction / methods*
  • RNA, Ribosomal / genetics
  • Reproducibility of Results
  • Sensitivity and Specificity


  • DNA, Protozoan
  • DNA, Ribosomal
  • RNA, Ribosomal