Phylogenetic neighbour-joining tree showing the relationship of the sigma-class GSTs of <i>Onchocerca ochengi</i> to similar enzymes of nematodes, mammals and insects.

<p>Numbers shown alongside branches are bootstrap values of 1,000 replications. The key for protein sequence accession numbers and organisms displayed in the tree is as follows: <u>Nematodes</u>: Oo_GST_t09064, Oo_GST_t03844 and Oo_GST_t06414 glutathione transferase [<i>Onchocerca ochengi</i>]; Ov_GST_1b AAG44696.1 glutathione <i>S</i>-transferase Ia [<i>Onchocerca volvulus</i>]; Ov _GST_1a AAG44695.1 glutathione <i>S</i>-transferase Ia [<i>Onchocerca volvulus</i>]; Ll_GST XP_003139665.1 hypothetical protein LOAG_04080 [<i>Loa loa</i>]; Bm_GST_4 XP_001901855.1 glutathione <i>S</i>-transferase 4 [<i>Brugia malayi</i>]; As_GST_1 ERG83753.1 glutathione <i>S</i>-transferase 1 [<i>Ascaris suum</i>]; As_GST_4 ERG81431.1 glutathione s-transferase 4 [<i>Ascaris suum</i>]; Ce_GST-11 NP_508625.1 protein GST-11 [<i>Caenorhabditis elegans</i>]; Ce_GST-36 NP_509652.2 protein GST-36 [<i>Caenorhabditis elegans</i>]. <u>Mammals:</u> Hs_PGD NP_055300.1 hematopoietic prostaglandin D synthase [<i>Homo sapiens</i>]; Bt_PGD_x1 XP_002688181.1 PREDICTED: hematopoietic prostaglandin D synthase isoform X1 [<i>Bos taurus</i>]; Rt_PGD NP_113832.1 hematopoietic prostaglandin D synthase [<i>Rattus norvegicus</i>]; Mm_PGD NP_062328.3 hematopoietic prostaglandin D synthase [<i>Mus musculus</i>]. <u>Insects:</u> Md_GST_ NP_001273827.1 glutathione <i>S</i>-transferase [<i>Musca domestica</i>]; Dm_GST_s1 NP_725653.1 glutathione <i>S</i>-transferase S1, isoform A [<i>Drosophila melanogaster</i>]; Ph_GST XP_002426887.1 glutathione <i>S</i>-transferase, putative [<i>Pediculus humanus corporis</i>]; Tc_GST XP_970714.1 PREDICTED: glutathione <i>S</i>-transferase [<i>Tribolium castaneum</i>]. The GSTs from <i>O. volvulus</i> and their closest relative in <i>O. ochengi</i> are shown in bold.</p

Alignment of partial gene sequences of glutathione <i>S</i>-transferases (GSTs) from <i>O. volvulus</i> (<i>OvGST1a</i>, <i>OvGST1b</i>) and <i>O. ochengi (OoGST1)</i> (A) and primer sets targeting <i>OvGST1a</i> (B).

<p>Primers are indicated by solid black arrows and dash arrows represent the binding regions of the loop forward (LFP) and loop back (LBP) primers respectively.</p

Sensitivity of LAMP and PCR methods for the detection of <i>O. volvulus</i> using ten-fold serial dilutions of <i>O. volvulus</i> genomic DNA ranging from 0.001–1.0 ng.

<p>Detection of LAMP product using turbidity (<b>A</b>) or hydroxy napthol blue (<b>B</b>). PCR amplification of a ∼200 bp product using LAMP primers F3 and B3 was obtained when <i>O. volvulus</i> genomic DNA was used (<b>C</b>). Molecular weight marker (MW) is indicated.</p

Diagrammatic view of the similarity of <i>Onchocerca</i> sigma-class GST gene models for <i>O. volvulus</i> GSTs 1a and 1b and the homologous <i>O. ochengi</i> sigma-class GST t09064.

<p>Gene models were aligned over the full-length sequence (total distance, 3,870 bp). Numbers associated with gene model exons (<i>I–VII shaded blocks</i>) and introns (<i>1–8 non-shaded blocks</i>) display the number of base-pairs within those sections over which the alignment is spaced. The three major differences between the genes (all insertions in <i>O. volvulus GST1a</i> intron 3) are highlighted in the diagram.</p

A comparison of LAMP and PCR methods to detect varying amounts of genomic DNA isolated from pools of black flies using different methods.

<p>A comparison of LAMP and PCR methods to detect varying amounts of genomic DNA isolated from pools of black flies using different methods.</p