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Clin Chem ; 52(2): 303-6, 2006 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-16339303

RESUMO

BACKGROUND: Malaria is one of the most important parasitic infections in humans. A sensitive diagnostic test for malaria that could be applied at the community level could be useful in programs to control the disease. The aim of the present work was to develop a simple, inexpensive molecular test for Plasmodium falciparum. METHODS: Blood was collected from controls (n = 100) and from patients diagnosed with falciparum malaria infection (n = 102), who were recruited to the study. Heat-treated blood samples were tested by a loop-mediated isothermal amplification (LAMP) assay for P. falciparum. Results were interpreted by a turbidity meter in real time or visually at the end of the assay. To evaluate the assay, DNA from these samples was purified and tested by PCR. Results from the LAMP and PCR assays were compared. RESULTS: The LAMP assay detected P. falciparum directly from heat-treated blood. The quantitative data from the assay correlated to the parasite counts obtained by blood-film microscopic analyses. When we used the PCR assay as the comparison method, the sensitivity and specificity of the LAMP assay were 95% and 99%, respectively. CONCLUSIONS: Unlike PCR, the LAMP assay does not require purified DNA for efficient DNA amplification, thereby reducing the cost and turnaround time for P. falciparum diagnosis. The assay requires only basic instruments, and assay positivity can be verified by visual inspection.


Assuntos
DNA de Protozoário/sangue , Temperatura Alta , Malária Falciparum/diagnóstico , Plasmodium falciparum/isolamento & purificação , Reação em Cadeia da Polimerase , Manejo de Espécimes/métodos , Animais , Sequência de Bases , Calibragem , DNA de Protozoário/genética , Humanos , Malária Falciparum/sangue , Dados de Sequência Molecular , Plasmodium falciparum/genética , Reação em Cadeia da Polimerase/economia , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/normas , Sensibilidade e Especificidade
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