Development and validation of a multiplexed-tandem qPCR tool for diagnostics of human soil-transmitted helminth infections

Stracke, K; Clarke, N; Awburn, CV; Vaz Nery, S; Khieu, V; Traub, RJ; Jex, AR

Author(s): Stracke, K; Clarke, N; Awburn, CV; Vaz Nery, S; Khieu, V; Traub, RJ; Jex, AR;
Details: Publication Year 2019-06-17,Volume 13,Issue #6,Page e0007363
Journal Title: PLoS Neglected Tropical Diseases
Publication Type: Journal Article
Abstract: Soil-transmitted helminths (STH) are a major cause of morbidity in tropical developing countries with a global infection prevalence of more than one billion people and disease burden of around 3.4 million disability adjusted life years. Infection prevalence directly correlates to inadequate sanitation, impoverished conditions and limited access to public health systems. Underestimation of infection prevalence using traditional microscopy-based diagnostic techniques is common, specifically in populations with access to benzimidazole mass treatment programs and a predominance of low intensity infections. In this study, we developed a multiplexed-tandem qPCR (MT-PCR) tool to identify and quantify STH eggs in stool samples. We have assessed this assay by measuring infection prevalence and intensity in field samples of two cohorts of participants from Timor-Leste and Cambodia, which were collected as part of earlier epidemiological studies. MT-PCR diagnostic parameters were compared to a previously published multiplexed qPCR for STH detection. The MT-PCR assay agreed strongly with qPCR data and showed a diagnostic specificity of 99.60-100.00% (sensitivity of 83.33-100.00%) compared to qPCR and kappa agreement exceeding 0.85 in all tests. In addition, the MT-PCR has the added advantage of distinguishing Ancylostoma spp. species, namely Ancylostoma duodenale and Ancylostoma ceylanicum. This semi-automated platform uses a standardized, manufactured reagent kit, shows excellent run-to-run consistency/repeatability and supports high-throughput detection and quantitation at a moderate cost.
Publisher: PLOS
Research Division(s): Population Health And Immunity
PubMed ID: 31206520
Publisher's Version: https://doi.org/10.1371/journal.pntd.0007363
Open Access at Publisher's Site: https://doi.org/10.1371/journal.pntd.0007363
NHMRC Grants: NHMRC/1126395,
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2019-06-20 10:18:11

Last Modified: 2019-06-20 12:21:32