Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, Australia.
Clinical Tropical Medicine Laboratory, QIMR Berghofer Medical Research Institute, Herston, Australia.
Am J Trop Med Hyg. 2018 Nov;99(5):1186-1193. doi: 10.4269/ajtmh.18-0276.
Hookworm disease caused by , , and affects half a billion people worldwide. The prevalence and intensity of infection of individual hookworm species are vital for assessing morbidity and generating targeted intervention programs for their control. The present study aims to evaluate a multiplex real-time quantitative PCR (qPCR) assay to determine the prevalence and egg intensity of all three hookworm species and compare this with standard microscopy and published genus-based conventional and real-time multiplex qPCRs. Performance of the diagnostic assays was evaluated using DNA extracted from 192 fecal samples collected as part of a soil-transmitted helminth (STH) survey in northern Cambodia. The prevalence of hookworms as detected by the multiplex hookworm qPCR of 84/192 (43.8%) was significantly higher than that using microscopy of 49/192 (25.5%). The hookworm multiplex qPCR showed very good agreement for the detection of both (Kappa 0.943) and spp. (Kappa 0.936) with a multiplex STH qPCR. A strong and moderate quantitative correlation between cycle threshold and eggs per gram (EPG) feces was obtained for the hookworm qPCR for seeded DNA egg extracts ( ≥ 0.9004) and naturally egg-infected individuals ( = 0.6848), respectively. The newly developed hookworm quantitative multiplex qPCR has the potential for application in anthelmintic efficacy trials and for monitoring the success of mass deworming programs targeting individual species of anthroponotic and zoonotic hookworms.
由 、 和 引起的钩虫病影响着全球半数人口。评估钩虫病的发病率和制定有针对性的防治方案，关键在于评估各钩虫种的流行程度和感染强度。本研究旨在评估一种多重实时定量 PCR (qPCR) 检测方法，以确定所有三种钩虫的流行程度和虫卵强度，并将其与标准显微镜检查和已发表的基于属的传统和实时多重 qPCR 进行比较。使用从柬埔寨北部土壤传播性蠕虫 (STH) 调查中收集的 192 份粪便样本中提取的 DNA 评估诊断检测的性能。使用多重钩虫 qPCR 检测到的钩虫流行率为 84/192（43.8%），明显高于显微镜检查的 49/192（25.5%）。多重钩虫 qPCR 对 和 种的检测具有很好的一致性（Kappa 0.943），与多重 STH qPCR 一致。对于种子 DNA 卵提取物（ ≥ 0.9004）和自然卵感染个体（ = 0.6848），钩虫 qPCR 的循环阈值和粪便每克卵（EPG）之间存在很强和中度的定量相关性。新开发的钩虫定量多重 qPCR 具有在驱虫疗效试验中应用的潜力，并可用于监测针对人源和动物源钩虫的大规模驱虫计划的成功。
