Plasmodium vivax: Reverse transcriptase real-time PCR for gametocyte detection and quantitation in clinical samples
Contribuinte(s) |
UNIVERSIDADE DE SÃO PAULO |
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Data(s) |
06/11/2013
06/11/2013
2012
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Resumo |
The proportion of Plasmodium vivax-infected subjects that carry mature gametocytes, and thus are potentially infectious, remains poorly characterized in endemic settings. Here, we describe a quantitative reverse transcriptase (RI) real-time PCR (qRT-PCR) that targets transcripts of the mature gametocyte-specific pvs25 gene. We found mature gametocytes in 42 of 44 (95.4%) P. vivax infections diagnosed during an ongoing cohort study in northwestern Brazil. SYBR green qRT-PCR was more sensitive than a conventional RT-PCR that targets the same gene. Molecular detection of gametocytes failed, however, when dried bloodspots were used for RNA isolation and complementary DNA synthesis. Estimating the number of pvs25 gene transcripts allowed for examining the potential infectiousness of gametocyte carriers in a quantitative way. We found that most (61.9%) gametocyte carriers were either asymptomatic or had subpatent parasitemias and would have been missed by routine malaria control strategies. However, potentially undiagnosed gametocyte carriers usually had low-density infections and contributed a small fraction (up to 4%) to the overall gametocyte burden in the community. Further studies are required to determine the relative contribution to malaria transmission of long-lasting but low-density gametocytemias in asymptomatic carriers that are left undiagnosed and untreated. (C) 2012 Elsevier Inc. All rights reserved. National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health of USA National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health of USA [RO1 AI 075416, U19 AI089681] Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP), Brazil Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP), Brazil [2009/52729-9] FAPESP FAPESP Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES), Brazil Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES), Brazil Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) of Brazil Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) of Brazil |
Identificador |
EXPERIMENTAL PARASITOLOGY, SAN DIEGO, v. 132, n. 3, supl. 1, Part 3, pp. 348-354, NOV, 2012 0014-4894 http://www.producao.usp.br/handle/BDPI/42481 10.1016/j.exppara.2012.08.010 |
Idioma(s) |
eng |
Publicador |
ACADEMIC PRESS INC ELSEVIER SCIENCE SAN DIEGO |
Relação |
EXPERIMENTAL PARASITOLOGY |
Direitos |
closedAccess Copyright ACADEMIC PRESS INC ELSEVIER SCIENCE |
Palavras-Chave | #PLASMODIUM VIVAX #GAMETOCYTES #DIAGNOSIS #REAL-TIME PCR #MALARIA CONTROL #FALCIPARUM #INFECTION #CARRIAGE #AMAZON #BRAZIL #ASSAY #RNA #PARASITOLOGY |
Tipo |
article original article publishedVersion |