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

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

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

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

Comparison of Malaria HTP-LAMP results from 699 DBS and WB samples against the equivalent gold standard nPCR results from 2011.

<p>Comparison of Malaria HTP-LAMP results from 699 DBS and WB samples against the equivalent gold standard nPCR results from 2011.</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 the study.

<p>The results of the two index tests, DBS-HTP-LAMP and WB-HTP-LAMP, against the gold standard nPCR results are summarized. Discrepancies between the gold standard and the index tests are highlighted. Abbreviation: EDTA ethylenediaminetetraacetic acid; PCR polymerase chain reaction; HTP-LAMP dried blood spot high throughput loop mediated isothermal amplification; DBS dried blood spot; WB whole blood.</p

The transportable hard-cover case containing the high throughput LAMP set-up.

<p>The transportable hard-cover case containing the high throughput LAMP set-up.</p

Layout of the contents of the high throughput sample processing kit on the bench.

<p>The contents of the sample processing kit which includes a LYSIS plate, PURE plate, EXTRACT plate, sample record sheet, lysis fluid, individual foam plugs and instruction manual.</p