Comparison between microscopy, conventional semi-nested PCR and real-time PCR-HRM assays (N = 634).

a<p>Details have been published elsewhere <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041996#pone.0041996-Ngui1" target="_blank">[2]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041996#pone.0041996-Ngui2" target="_blank">[34]</a>.</p>b<p>Sensitivity: 84.1%; Specificity: 100% (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041996#pone.0041996.s001" target="_blank">Figure S1</a>).</p>c<p>Sensitivity: 100%; Specificity: 100% (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041996#pone.0041996.s001" target="_blank">Figure S1</a>).</p

Hookworm species detected via both conventional semi-nested PCR and real-time PCR-HRM assays (N = 58).

a<p>Details have been published elsewhere <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041996#pone.0041996-Ngui1" target="_blank">[2]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041996#pone.0041996-Ngui2" target="_blank">[34]</a>.</p

Representative profiles of the melting curves (aligned melt curves) of ITS-2 amplicons for <i>Necator americanus</i> (black), <i>Ancylostoma duodenale</i> (blue), <i>A. ceylanicum</i> (red), <i>A. caninum</i> (green) and <i>A. braziliense</i> (purple).

<p>Fluorescence is plotted against degrees Celsius (°C).</p

Representative profiles of the melting curves (difference plot curves) of ITS-2 amplicons for <i>Necator americanus</i> (black), <i>Ancylostoma duodenale</i> (blue), <i>A. ceylanicum</i> (red), <i>A. caninum</i> (green) and <i>A. braziliense</i> (purple).

<p>Representative profiles of the melting curves (difference plot curves) of ITS-2 amplicons for <i>Necator americanus</i> (black), <i>Ancylostoma duodenale</i> (blue), <i>A. ceylanicum</i> (red), <i>A. caninum</i> (green) and <i>A. braziliense</i> (purple).</p

Results achieved by real-time PCR coupled HRM analysis of ITS-2 amplicon from control genomic DNA for hookworm.

<p>Results achieved by real-time PCR coupled HRM analysis of ITS-2 amplicon from control genomic DNA for hookworm.</p

Antibody Prevalence and Factors Associated with Exposure to <i>Orientia tsutsugamushi</i> in Different Aboriginal Subgroups in West Malaysia

<div><p>Background</p><p>Limited data is available on the current status of scrub typhus infection in the aboriginal population in Malaysia. This study was aimed to provide recent data on the degree of exposure of 280 individuals from seven aboriginal subgroups to <i>Orientia tsutsugamushi</i> (causative agent of scrub typhus) in West Malaysia. The environment, socioeconomic and behavioural risk factors associated with the disease were also investigated.</p><p>Methods/Findings</p><p>The antibody prevalence to <i>O. tsutsugamushi</i> ranged from 0 to 36.4% in seven subgroups, with high prevalence rates noted in subgroups involved in agricultural activity and the lowest prevalence rates noted in subgroups whose main occupations were associated to fishing. Univariate analysis indicated populations with age above 18 years (OR = 1.15, 95% CI = 1.02–1.30, <i>P</i> = 0.015), working (OR = 1.99, 95% CI = 1.01–3.92, <i>P</i> = 0.044), working at agriculture area (OR = 1.18, 95% CI = 0.98–1.42, <i>P</i> = 0.031), receiving household income less than US$166.7 (RM500) per month (OR = 2.43, 95$ 166.7 (RM500) per month and having close contact with animal pets are 3.6 times (95% CI = 1.81–7.03, <i>P</i><0.001), 1.3 times (95% CI = 1.14–1.64, <i>P</i> = 0.002) and 1.2 times (95% CI = 1.05–1.06, <i>P</i> = 0.006) more likely to have exposure to <i>O. tsutsugamushi</i>, respectively.</p><p>Conclusion</p><p>The present study indicates that scrub typhus is still an important disease in the aboriginal population in Malaysia. Awareness about the disease and education on the preventive measures are important in reducing the risk of acquiring scrub typhus in the population studied.</p></div

Demographic and baseline characteristics of the surveyed population (n = 280).

<p>Demographic and baseline characteristics of the surveyed population (n = 280).</p

Prevalence of IgG antibodies to <i>O. tsutsugamushi</i> among the surveyed population (n = 280).

<p>Prevalence of IgG antibodies to <i>O. tsutsugamushi</i> among the surveyed population (n = 280).</p

Potential associated factors associated with hookworm infection by microscopy in human (Univariate analysis, N = 634).

<p>Reference group marked as OR = 1; 95% CI: 95% Confidence interval; Significant association.</p><p>(<i>p</i><0.05).</p><p>*Variables were confirmed by multivariate analysis as significant predictors of hookworm infection.</p

Hookworm species detected from human and animal samples as determined by PCR assays.

<p>*<i>A. caninum</i> were found in dogs;</p><p>**<i>A. ceylanicum</i> were found in both cats and dogs;</p><p>***<i>A. braziliense</i> was found in cats.</p