Malaria diagnosis: standardization of a polymerase chain reaction for the detection of Plasmodium falciparum parasites in individuals with low-grade parasitemia
In Brazil, no study has been done concerning the detection of malaria parasites by polymerase chain reaction (PCR) related to the diagnosis of Plasmodium falciparum malaria. In the present report we describe a highly sensitive methodology for malaria diagnosis using a nested PCR method based on...
| Autor principal: | Zalis, M. G |
|---|---|
| Outros Autores: | Cruz, M. F. Ferreira da, Guedes, H. C. Balthazar, Banic, D. M, Alecrim, Wilson, Souza, Jos? Maria de, Druilhe, P, Daniel-Ribeiro, Claudio Tadeu |
| Grau: | Artigo |
| Idioma: | eng |
| Publicado em: |
Springer Verlag
2019
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| Assuntos: | |
| Acesso em linha: |
http://patua.iec.gov.br//handle/iec/3566 |
| Resumo: |
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In Brazil, no study has been done concerning
the detection of malaria parasites by polymerase chain
reaction (PCR) related to the diagnosis of Plasmodium
falciparum malaria. In the present report we describe a
highly sensitive methodology for malaria diagnosis using
a nested PCR method based on amplification of the p126
P. falciparum gene detected by simple ethidium bromide
staining. The P. falciparum Palo Alto strain (culture samples) was serially diluted in blood from an uninfected
donor to a final level of parasitemia corresponding to
10?8% and was processed for PCR amplification. In each
of these dilutions a parasitological examination was performed to compare the sensitivity with that of PCR amplification. Blood samples (field samples) were obtained
from 51 malarious patients with positive thick blood
smears (TBS) who were living in endemic regions of the
Brazilian Amazon. They corresponded to 42 P. falciparum and 9 P. vivax cases, with parasitemia levels ranging
from only 16 to 20,200 parasites/?l for P. falciparum disease and from 114 to 11,000 parasites/?l for P. vivax malaria. We demonstrate that the use of nested PCR allows
the detection of 0.005 parasites/?l without the use of radioactive material. The use of a 1-ml sample volume and
the organic DNA extraction method should be suitable in blood banks and for the evaluation of patients during and
after drug treatment. |