Plasmodium vivax: parasitemia determination by real-time quantitative PCR in Aotus monkeys
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 parasite...
- Autores:
- Tipo de recurso:
- Fecha de publicación:
- 2002
- Institución:
- Universidad del Rosario
- Repositorio:
- Repositorio EdocUR - U. Rosario
- Idioma:
- eng
- OAI Identifier:
- oai:repository.urosario.edu.co:10336/28473
- Acceso en línea:
- https://doi.org/10.1016/S0014-4894(02)00010-3
https://repository.urosario.edu.co/handle/10336/28473
- Palabra clave:
- Plasmodium vivax
Aotus nancymaae
Real-time quantitative
Green I
Parasitemia
Small subunit ribosomal RNA
SYBR
PCR
- Rights
- License
- Restringido (Acceso a grupos específicos)
Summary: | 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. |
---|