Supplementary dataset.

All data that support the findings of this study are available as a supplementary document uploaded to the journal’s website. (XLSX)</p

DiaxxoPCR and PlasmoPod setup for rapid molecular malaria detection.

A) The DiaxxoPCR instrument is a pyramid-shaped stand-alone device for rapid qPCR cycling and fluorescence acquisition. (B) The PlasmoPod assay is based on a single-use cartridge which is pre-loaded with all qPCR reagents in dried form. (C) The PlasmoPod laboratory setup consisting of the diaxxoPCR device, a heatblock, a pipette and PlasmoPod cartridges. (D) Finger prick blood is sampled and stored as dried blood spots (DBS). (E) The setup for rapid NA extraction from DBS. (F) The NA are amplified and signals analysed using the diaxxoPCR device and its integrated analysis software. Images A and B are republished from diaxxo AG under a CC BY license, with permission from Dr Michele Gregorini (who is an author of this manuscript), original copyright [diaxxo AG, 2020–2022]. The granted permission to the manuscript has been uploaded. Partially created with Biorender.com.</p

Parasitological and demographic characteristics of the study population selected for evaluation of PlasmoPod from Central African Republic.

Parasitological and demographic characteristics of the study population selected for evaluation of PlasmoPod from Central African Republic.</p

Molecular analysis of <i>Plasmodium</i> spp. parasites identified in the CAR dataset.

Three different molecular assays were used to (A) screen for Plasmodium spp. parasites, (B) identify Plasmodium spp. species and (C) detect pfhrp2/3 gene deletion. Each child is represented in a column stratified according to malaria infection status. Green colors represent negative measurements for the respective qPCR assay, while grey colors were chosen for tests which were not conducted. All tests were run on the Bio-Rad CXF96 qPCR instrument. (TIF)</p

Analysis of asymptomatic malaria cohort.

(A) Correlation of of Cq valuesvales obtained from reference RT-qPCR run on the Biorad CFX96 instrumentintrument and PlasmoPod run on diaxxoPCR. Samples negative for PlasmoPod were assigned a Cq value of -1. (B) Detection probability for PlasmoPod modelled based on reference Cq values. The grey area represents the 95% confidence interval. (C) Detection rate of PlasmoPod stratified by age group. (D) Cq values stratified by age group.</p

Diagnostic performance of RDT (PfHRP2/panLDH), TBS microscopy and PlasmoPod compared to the gold standard RT-qPCR assay run on the BioRad CFX96 qPCR instrument.

Diagnostic performance of RDT (PfHRP2/panLDH), TBS microscopy and PlasmoPod compared to the gold standard RT-qPCR assay run on the BioRad CFX96 qPCR instrument.</p

Analytical performance evaluation of PlasmoPod.

(A) DiaxxoPCR derived delta fluorescence and maximum slope after amplification with nucleic acid from different pathogens. Lines indicate the delta fluorescence positivity cutoff (vertical) and maximum slope positivity cutoff (horizontal). (B) Cq value of PlasmoPod versus dilutions of NAs extracted from culture-derived ring-stage synchronized P. falciparum parasites. Sample without amplification are colored in red.</p

Quantification of <i>P</i>. <i>falciparum</i> parasites using PlasmoPod.

(A) Correlation of Cq values derived from PlasmoPod and reference (RT)-qPCR assays based on Plasmodium spp. 18S ribosomal DNA and RNA (left panel) and the P. falciparum ribonucleotide reductase R2_e2 assays (right panel). (B) Correlation between PlasmoPod and thick blood smear microscopy for quantification of P. falciparum parasites. The grey color represents the 95% confidence interval.</p

Clinical and parasitological characterization of study population.

(A) Correlation of parasite density assessed by the PfRNR2 assay and thick blood smear microscopy. (B) Modelled distribution of parasite densities measured by PfRNR2 assay and highlighted for high- and moderate-density infections. (C) Hematological parameters, including red blood cell counts (RBC), Hemoglobin levels (Hb), Thrombocyte counts (PLT) and white blood cell counts (WBC) compared between malaria negative children and children with high- and moderate-density malaria infections.</p