Please use this identifier to cite or link to this item: http://www.arca.fiocruz.br/handle/icict/9403
Title: Submicroscopic malaria parasite carriage: how reproducible are polymerase chain reaction-based methods?
Authors: Costa, Daniela Camargos
Madureira, Ana Paula
Amaral, Lara Cotta
Sanchez, Bruno Antônio Marinho
Gomes, Luciano Teixeira
Fontes, Cor Jésus Fernandes
Limongi, Jean Ezequiel
Brito, Cristiana Ferreira Alves de
Carvalho, Luzia Helena
Affilliation: Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil
Universidade Federal de São João Del Rei. Departamento de Bioengenharia. São João Del Rey, MG, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil
Universidade Federal de Mato Grosso. Cuiabá, MT, Brasil
Universidade Federal de Mato Grosso. Cuiabá, MT, Brasil
Universidade Federal de Mato Grosso. Cuiabá, MT, Brasil
Centro de Controle de Zoonoses de Uberlândia. Uberlândia, MG, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil
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.
Keywords: malaria
PCR
reproducibility
submicroscopic infection
Issue Date: 2014
Publisher: Fundação Oswaldo Cruz
Citation: COSTA, Daniela Camargos et al. Submicroscopic malaria parasite carriage: how reproducible are polymerase chain reaction-based methods?. Mem. Inst. Oswaldo Cruz, Rio de Janeiro , v. 109, n. 1, Feb. 2014.
DOI: http://dx.doi.org/10.1590/0074-0276140102
ISSN: 0074-0276
Copyright: open access
Appears in Collections:IRR - Artigos de Periódicos

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