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

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

Resultado de la investigación: Contribución a una revistaArtículorevisión exhaustiva

16 Citas (Scopus)

Resumen

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.

Idioma originalInglés estadounidense
Páginas (desde-hasta)131-134
Número de páginas4
PublicaciónExperimental Parasitology
Volumen100
N.º2
DOI
EstadoPublicada - feb. 1 2002
Publicado de forma externa

Áreas temáticas de ASJC Scopus

  • Parasitología
  • Inmunología
  • Enfermedades infecciosas

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