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

Juan Carlos Polanco, Josefa A. Antonia Rodrı́guez, Vladimir Corredor, Manuel Alfonso Patarroyo

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)131-134
Number of pages4
JournalExperimental Parasitology
Volume100
Issue number2
DOIs
StatePublished - Feb 1 2002
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Parasitology
  • Immunology
  • Infectious Diseases

Fingerprint Dive into the research topics of 'Plasmodium vivax: Parasitemia determination by real-time quantitative PCR in Aotus monkeys'. Together they form a unique fingerprint.

Cite this