Submicroscopic malaria parasite carriage: how reproducible are polymerase chain reaction-based methods?
Mem. Inst. Oswaldo Cruz
;
109(1): 21-28, 02/2014. tab, graf
Artigo
em Inglês
| LILACS
| ID: lil-703648
ABSTRACT
The polymerase chain reaction (PCR)-based methods for the diagnosis of malaria infection are expected to accurately identify submicroscopic parasite carriers. Although a significant number of PCR protocols have been described, few studies have addressed the performance of PCR amplification in cases of field samples with submicroscopic malaria infection. Here, the reproducibility of two well-established PCR protocols (nested-PCR and real-time PCR for the Plasmodium 18 small subunit rRNA gene) were evaluated in a panel of 34 blood field samples from individuals that are potential reservoirs of malaria infection, but were negative for malaria by optical microscopy. Regardless of the PCR protocol, a large variation between the PCR replicates was observed, leading to alternating positive and negative results in 38% (13 out of 34) of the samples. These findings were quite different from those obtained from the microscopy-positive patients or the unexposed individuals; the diagnosis of these individuals could be confirmed based on the high reproducibility and specificity of the PCR-based protocols. The limitation of PCR amplification was restricted to the field samples with very low levels of parasitaemia because titrations of the DNA templates were able to detect < 3 parasites/µL in the blood. In conclusion, conventional PCR protocols require careful interpretation in cases of submicroscopic malaria infection, as inconsistent and false-negative results can occur.
Texto completo:
DisponíveL
Índice:
LILACS (Américas)
Assunto principal:
Plasmodium
/
Portador Sadio
/
Reação em Cadeia da Polimerase
/
DNA de Protozoário
/
Malária
Tipo de estudo:
Estudo diagnóstico
/
Guia de Prática Clínica
/
Estudo prognóstico
Limite:
Adolescente
/
Adulto
/
Idoso
/
Aged80
/
Feminino
/
Humanos
/
Masculino
Idioma:
Inglês
Revista:
Mem. Inst. Oswaldo Cruz
Assunto da revista:
Medicina Tropical
/
Parasitologia
Ano de publicação:
2014
Tipo de documento:
Artigo
País de afiliação:
Brasil
Instituição/País de afiliação:
Fiocruz/BR
Similares
MEDLINE
...
LILACS
LIS