A morphology-based method for the diagnosis of red blood cells parasitized by Plasmodium malariae and Plasmodium ovale

Karimi, Alireza; Navidbakhsh, Mahdi; Haghi, Afsaneh Motevalli; Faghihi, Shahab

doi:10.3109/00365548.2014.880186

Cited by 2 publications

(1 citation statement)

References 28 publications

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“…For example, our initial attempts on P. ovale clinical samples and on in vitro P. falciparum culture containing more mature asexual stages (trophozoite and schizont) indicated highly variable results regarding the capacity of these other species and stages to pass through the 5 μm filter we used (data not shown). Considering that both the size and deformability of the host RBC vary according to the infecting Plasmodium species [31,32] and stage maturity, a 5 μm filter might not be suitable for other Plasmodium species infecting humans.…”

Section: Discussionmentioning

confidence: 99%

5WBF: A low-cost and straightforward whole blood filtration method suitable for whole-genome sequencing of Plasmodium falciparum clinical isolates

Coppée

Mama

Sarrasin

et al. 2021

Preprint

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Background: Whole-genome sequencing (WGS) is becoming increasingly helpful to assist malaria control programs. A major drawback of this approach is the large amount of human DNA compared to parasite DNA extracted from unprocessed whole blood. As red blood cells (RBCs) have a diameter of about 7-8 μm and exhibit some deformability, we hypothesized that cheap and commercially available 5 μm filters might retain leukocytes but much less of Plasmodium falciparum-infected RBCs. This study aimed to test the hypothesis that such a filtration method, named 5WBF (for 5 μm Whole Blood Filtration), may provide highly enriched parasite material suitable for P. falciparum WGS. Methods: Whole blood was collected from five patients experiencing a P. falciparum malaria episode (ring-stage parasitemia range: 0.04-5.5%) and from mock samples obtained by mixing synchronized, ring-stage cultured P. falciparum 3D7 parasites with uninfected human whole blood (final parasitemia range: 0.02-1.1%). These whole blood samples (50 to 400 μL) were diluted in RPMI 1640 medium or PBS 1X buffer and filtered with syringes connected to a 5 μm commercial filter. DNA was extracted from filtered and unfiltered counterpart blood samples using a commercial kit. The 5WBF method was evaluated on the ratios of parasite:human DNA assessed by qPCR and by sequencing depth and percentages of coverage from WGS data (Illumina NextSeq 500). As a comparison, we also applied to the same unprocessed whole blood samples the selective whole-genome amplification (sWGA) method which does not rely on blood filtration. Results: After applying 5WBF, qPCR indicated an average of 2-fold loss in the amount of parasite template DNA (Pf ARN18S gene) and from 4,096- to 65,536-fold loss of human template DNA (human β actin gene). WGS analyses revealed that > 95% of the nuclear genome and the entire whole organellar genomes were covered at ≥ 10x depth for all samples tested. In sWGA counterparts, none of the organellar genomes were covered, and from 47.7 to 82.1% of the nuclear genome was covered at ≥ 10x depth depending on parasitemia. Sequence reads were homogeneously distributed across gene sequences for 5WBF-treated samples (n = 5,460 genes; mean coverage: 91x; median coverage: 93x; 5th percentile: 70x; 95th percentile: 103x), allowing the identification of gene copy number variations such as for gch1. This later analysis was not possible for sWGA-treated samples, as we observed a much more heterogeneous distribution of reads among gene sequences (mean coverage: 80x; median coverage: 51x; 5th percentile: 7x; 95th percentile: 245x. Conclusions: The novel 5WBF leucodepletion method is simple to implement and based on commercially available, standardized, 5 μm filters which cost from 1.0 to 1.7€ per unit, depending on suppliers. 5WBF permits extensive genome-wide analysis of P. falciparum DNA from minute amounts of whole blood even with parasitemias as low as 0.02%.

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Section: Discussionmentioning

confidence: 99%

5WBF: A low-cost and straightforward whole blood filtration method suitable for whole-genome sequencing of Plasmodium falciparum clinical isolates

Coppée

Mama

Sarrasin

et al. 2021

Preprint

View full text Add to dashboard Cite

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Advances in Malaria Diagnostic Methods in Resource-Limited Settings: A Systematic Review

Yalley,

Ocran,

Cobbinah

et al. 2024

TropicalMed

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Malaria continues to pose a health challenge globally, and its elimination has remained a major topic of public health discussions. A key factor in eliminating malaria is the early and accurate detection of the parasite, especially in asymptomatic individuals, and so the importance of enhanced diagnostic methods cannot be overemphasized. This paper reviewed the advances in malaria diagnostic tools and detection methods over recent years. The use of these advanced diagnostics in lower and lower-middle-income countries as compared to advanced economies has been highlighted. Scientific databases such as Google Scholar, PUBMED, and Multidisciplinary Digital Publishing Institute (MDPI), among others, were reviewed. The findings suggest important advancements in malaria detection, ranging from the use of rapid diagnostic tests (RDTs) and molecular-based technologies to advanced non-invasive detection methods and computerized technologies. Molecular tests, RDTs, and computerized tests were also seen to be in use in resource-limited settings. In all, only twenty-one out of a total of eighty (26%) low and lower-middle-income countries showed evidence of the use of modern malaria diagnostic methods. It is imperative for governments and other agencies to direct efforts toward malaria research to upscale progress towards malaria elimination globally, especially in endemic regions, which usually happen to be resource-limited regions.

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A morphology-based method for the diagnosis of red blood cells parasitized by Plasmodium malariae and Plasmodium ovale

Cited by 2 publications

References 28 publications

5WBF: A low-cost and straightforward whole blood filtration method suitable for whole-genome sequencing of Plasmodium falciparum clinical isolates

5WBF: A low-cost and straightforward whole blood filtration method suitable for whole-genome sequencing of Plasmodium falciparum clinical isolates

Advances in Malaria Diagnostic Methods in Resource-Limited Settings: A Systematic Review

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