Additional file 1: of A simple, high-throughput, colourimetric, field applicable loop-mediated isothermal amplification (HtLAMP) assay for malaria elimination

Features of asymptomatic Ghanaian children samples (n=25). Table outlines features of the 25 samples chosen from a set of samples from asymptomatic school children in Kumasi, Ghana

DENV-2 phylogenetic tree based on partial M and E gene nucleotide sequences depicting the relationship between the virus acquired by the patient and that used in laboratory experiments (highlighted).

<p>These are compared with other recent DENV-2 isolates such as Townsville 1993 that were imported or resulted in outbreaks in Queensland. Also shown are the genotypes of DENV-2 and representative strains from each grouping.</p

HtLAMP colour change associated with hydroxynaphtholblue (HNB).

<p><i>Left</i> clear, purple colour = negative and <i>right</i> cloudy, blue colour = positive.</p

Analytical sensitivity of HtLAMP-Pv on microscopy-determined whole blood <i>P</i>. <i>vivax</i> dilution series.

<p>The limit of detection HtLAMP-Pv is 2 parasites/ μL, performed in duplicate and where both duplicates were positive.</p

Sensitivity and specificity of HtLAMP-Pv for <i>P</i>. <i>vivax</i> in symptomatic patients in Sabah with <i>P</i>. <i>falciparum</i>, <i>P</i>. <i>vivax</i> and <i>P</i>. <i>knowlesi</i>.

<p>149 filter paper samples were tested by HtLAMP-Pv; 4 samples were excluded due to a lack of PCR and microscopy data. Among the 145 samples, PCR confirmation of species was as follows: n = 64 <i>P</i>. <i>vivax</i> n = 56 <i>P</i>. <i>knowlesi</i>, n = 17 <i>P</i>. <i>falciparum</i>, n = 7 <i>P</i>. <i>malariae</i> and n = 1 mixed <i>P</i>. <i>vivax/P</i>. <i>knowlesi</i> infection.</p

<i>P</i>. <i>vivax</i> VIV2 HtLAMP primer sequences (5’ → 3’).

<p><i>P</i>. <i>vivax</i> VIV2 HtLAMP primer sequences (5’ → 3’).</p

Primers used to amplify Pv mdr, aldolase and cox 1 (*Source [36]).

<p>Primers used to amplify Pv mdr, aldolase and cox 1 (*Source [<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004443#pntd.0004443.ref036" target="_blank">36</a>]).</p

Comparing microscopy and RDT with the limit of detection of HtLAMP-Pv using filter paper and whole blood at 10 μL and 50 μL volumes.

<p>Comparing microscopy and RDT with the limit of detection of HtLAMP-Pv using filter paper and whole blood at 10 μL and 50 μL volumes.</p

Estimated <i>Plasmodium vivax</i> cox1 copy number based on comparison with two single copy genes, <i>pv aldolase1</i> and <i>pvmdr1</i>.

<p>Estimated <i>Plasmodium vivax</i> cox1 copy number based on comparison with two single copy genes, <i>pv aldolase1</i> and <i>pvmdr1</i>.</p

Analytical sensitivity of HtLAMP-Pv, compared with other published <i>P</i>. <i>vivax</i> LAMP primers, using a DNA dilution series of clinical sample with qPCR confirmed parasitemia.

<p>Each sample was tested in duplicate in the HtLAMP platform using each of the three <i>P</i>. <i>vivax</i> LAMP primer sets. The dilution at which both duplicates were positive was the limit of detection for each of the primer sets in the HtLAMP platform (Pos* indicates dilutions at which only one of the duplicates was positive). HtLAMP-Pv was able to detect 1.4 parasites/ μL compared with previously published <i>P</i>. <i>vivax</i> LAMP primers.</p