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Malaria PCR Detection in Cambodian Low-Transmission Settings: Dried Blood Spots versus Venous Blood Samples
Authors:Lydie Canier  Nimol Khim  Saorin Kim  Rotha Eam  Chanra Khean  Kaknika Loch  Malen Ken  Pieter Pannus  Philippe Bosman  Jorgen Stassijns  Fabienne Nackers  SweetC Alipon  Meng Chuor Char  Nguon Chea  William Etienne  Martin De Smet  Jean-Marie Kindermans  Didier Ménard
Affiliation:Institut Pasteur du Cambodge, Phnom Penh, Cambodia; Médecins Sans Frontières, Brussels, Belgium; Epicentre, Paris, France; Médecins Sans Frontières, Tuol Svay Prey I, Chamkarmon, Phnom Penh, Cambodia; National Center for Parasitology, Entomology and Malaria Control, Phnom Penh, Cambodia
Abstract:In the context of malaria elimination, novel strategies for detecting very low malaria parasite densities in asymptomatic individuals are needed. One of the major limitations of the malaria parasite detection methods is the volume of blood samples being analyzed. The objective of the study was to compare the diagnostic accuracy of a malaria polymerase chain reaction assay, from dried blood spots (DBS, 5 μL) and different volumes of venous blood (50 μL, 200 μL, and 1 mL). The limit of detection of the polymerase chain reaction assay, using calibrated Plasmodium falciparum blood dilutions, showed that venous blood samples (50 μL, 200 μL, 1 mL) combined with Qiagen extraction methods gave a similar threshold of 100 parasites/mL, ∼100-fold lower than 5 μL DBS/Instagene method. On a set of 521 field samples, collected in two different transmission areas in northern Cambodia, no significant difference in the proportion of parasite carriers, regardless of the methods used was found. The 5 μL DBS method missed 27% of the samples detected by the 1 mL venous blood method, but most of the missed parasites carriers were infected by Plasmodium vivax (84%). The remaining missed P. falciparum parasite carriers (N = 3) were only detected in high-transmission areas.
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