Asymptomatic individuals with discordant results.

<p>*determined by quantitative PCR, Pf = <i>Plasmodium falciparum</i> Pm = <i>Plasmodium malariae</i>,</p><p>P/µL = parasites/microliter, + positive, − negative.</p><p>Outcome summary of asymptomatic individuals with discordant results between.</p><p>Pan and/or Pf-LAMP and nested PCR using Chelex extracted DNA.</p

Limit of detection of LAMP reactions using the boil and spin DNA extraction method.

Representative amplifications experiments using serial dilutions of samples of known parasitaemia (quantified using the WHO International Standard as calibrator).Quantification was achieved by extracting DNA from a parasite dilution series made in whole blood using a Qiacube and DNA blood mini kit (Qiagen, De Hilden). Extracted DNA was then quantified using the Shokoples real time PCR [28] against a within run standard curve created from DNA extracted from the WHO international standard using the same methodology. Parasite count per microLitre was calculated given an initial parasite burden of 10% in the WHO standard and an assumed red cell count of 5 x106 red cells per microLitre. Mean time to turbidity in minutes as follows: 10,000 p/μL = 15.8; 1,000 p/μL = 15.9; 100 p/μL = 16.7; 10 p/μL = 18.2; 1 p/μL = 24.5.</p

The contents of high throughput LAMP set-up packed into a transportable hard-cover case.

The contents of high throughput LAMP set-up packed into a transportable hard-cover case.</p

Sensitivities, specificities, positive and negative predictive values and kappa analysis.

<p><i>P.f.</i>* = <i>P. falciparum</i> mono infections (n = 33), P.m.** = <i>P.malariae</i> mono infections (n = 13), P.f.*** <i>P. falciparum</i> mono and mixed.</p><p>infections (n = 41), CI = confidence interval, PPV = positive predictive value, NPV = negative predictive value.</p><p>Sensitivities, specificities, positive and negative predictive values and kappa analysis for detection of malaria DNA from fever patients and asymptomatic individuals with Pan and Pf-LAMP versus gold standard (real- time PCR corrected Cytochrome b nested PCR).</p

Fever patient samples with discordant results.

<p>*Determined by blood smear microscopy, RDT = rapid diagnostic test + positive, − negative.</p><p>Outcome summery of fever patient samples with discordant results between Pan and/or Pf-LAMP and nested PCR using ABI extracted DNA and after DNA re-extraction with Chelex.</p

Limit of detection of LAMP reactions using the high throughput sample processing system.

<p>Representative amplifications experiments using serial dilutions of samples of known parasitaemia (quantified using the WHO International Standard as calibrator). Mean time to turbidity in minutes as follows: 10,000 p/μL = 17.5; 1,000 p/μL = 18.5; 100 p/μL = 19.4; 10 p/μL = 21.3; 1 p/μL = 22.9.</p

Stratified analysis of parasitaemia comparing DBS and WB HTP-LAMP against nPCR with a cut-off at 2 p/μL.

<p>Stratified analysis of parasitaemia comparing DBS and WB HTP-LAMP against nPCR with a cut-off at 2 p/μL.</p

Flow chart of study. Reference standard

<p> = Cytochrome B nested PCR. <b>Gold standard</b> = Cytochrome B real-time PCR corrected nested PCR.</p

Baseline characteristics of fever patients and asymptomatic individuals.

<p>*determined by microscopy,</p><p>**determined by quantitative PCR,</p><p>ND = not done, p/µL = parasites/microliter, values in ( ) = range.</p

Layout of the high throughput set-up on the bench.

<p>The layout of the high throughput set-up which includes a shaker, hot block, vacuum manifold and pressure gauge.</p