PCR targeting Plasmodium mitochondrial genome of DNA extracted from dried blood on filter paper compared to whole blood.
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.
Department of Clinical Science, University of Bergen, Bergen, Norway. gro_strom@hotmail.com.
Gro E A
GE
Strøm
Sabrina
S
Moyo
Maulidi
M
Fataki
Nina
N
Langeland
Bjørn
B
Blomberg
D002648:Child; D003955:Diagnostic Tests, Routine / Q000379:methods*; D059788:Dried Blood Spot Testing / Q000379:methods*; D006801:Humans; D016778:Malaria, Falciparum / Q000097:blood / Q000175:diagnosis*; D008853:Microscopy / Q000379:methods*; D010963:Plasmodium falciparum / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D013636:Tanzania
D012337:RNA, Ribosomal, 18S