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Malar J. 2014 Apr 7;13:137. doi: 10.1186/1475-2875-13-137.

PCR targeting Plasmodium mitochondrial genome of DNA extracted from dried blood on filter paper compared to whole blood.

Author information

1
Department of Clinical Science, University of Bergen, Bergen, Norway. gro_strom@hotmail.com.

Abstract

BACKGROUND:

Monitoring mortality and morbidity attributable to malaria is paramount to achieve elimination of malaria. Diagnosis of malaria is challenging and PCR is a reliable method for identifying malaria with high sensitivity. However, blood specimen collection and transport can be challenging and obtaining dried blood spots (DBS) on filter paper by finger-prick may have advantages over collecting whole blood by venepuncture.

METHODS:

DBS and whole blood were collected from febrile children admitted at the general paediatric wards at a referral hospital in Dar es Salaam, Tanzania. DNA extracted from whole blood and from DBS was tested with a genus-specific PCR targeting the mitochondrial Plasmodium genome. Positive samples by PCR of DNA from whole blood were tested with species-specific PCR targeting the 18S rRNA locus, or sequencing if species-specific PCR was negative. Rapid diagnostic test (RDT) and thin blood smear microscopy was carried out on all patients where remnant whole blood and a blood slide, respectively, were available.

RESULTS:

Positivity of PCR was 24.5 (78/319) and 11.2% (52/442) by whole blood and DBS, respectively. All samples positive on DBS were also positive on Plasmodium falciparum species-specific PCR. All RDT positive cases were also positive by DBS PCR. All but three cases with positive blood slides were also positive by DBS.

CONCLUSIONS:

In this study, PCR for malaria mitochondrial DNA extracted from whole blood was more sensitive than from DBS. However, DBS are a practical alternative to whole blood and detected approximately the same number of cases as RDTs and, therefore, remain relevant for research purposes.

PMID:
24708551
PMCID:
PMC3983671
DOI:
10.1186/1475-2875-13-137
[Indexed for MEDLINE]
Free PMC Article
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