BACKGROUND Soil-transmitted helminths (STHs) are parasitic nematodes that inhabit the human intestine. They affect more than 1.5 billion people worldwide, causing physical and cognitive impairment in children. The global strategy to control STH infection includes periodic mass drug administration (MDA) based on the results of diagnostic testing among populations at risk, but the current microscopy method for detecting infection has diminished sensitivity as the intensity of infection decreases. Thus, improved diagnostic tools are needed to support decision-making for STH control programs. METHODOLOGY We developed a nucleic acid amplification test based on recombinase polymerase amplification (RPA) technology to detect STH in stool. We designed primers and probes for each of the four STH species, optimized the assay, and then verified its performance using clinical stool samples. PRINCIPAL FINDINGS Each RPA assay was as sensitive as a real-time polymerase chain reaction (PCR) assay in detecting copies of cloned target DNA sequences. The RPA assay amplified the target in DNA extracted from human stool samples that were positive for STH based on the Kato-Katz method, with no cross-reactivity of the non-target genomic DNA. When tested with clinical stool samples from patients with infections of light, moderate, and heavy intensity, the RPA assays demonstrated performance comparable to that of real-time PCR, with better results than Kato-Katz. This new rapid, sensitive and field-deployable method for detecting STH infections can help STH control programs achieve their goals. CONCLUSIONS Semi-quantitation of target by RPA assay is possible and is comparable to real-time PCR. With proper instrumentation, RPA assays can provide robust, semi-quantification of STH DNA targets as an alternative field-deployable indicator to counts of helminth eggs for assessing infection intensity.

中文翻译：

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使用基于重组酶聚合酶扩增的测定法对粪便中土壤传播的蠕虫感染进行灵敏和半定量检测。

背景土壤传播的蠕虫(STH) 是寄生于人体肠道的线虫。它们影响全球超过 15 亿人，导致儿童身体和认知障碍。控制 STH 感染的全球策略包括根据高危人群的诊断测试结果定期大量给药 (MDA)，但随着感染强度的降低，目前用于检测感染的显微镜方法已经降低了敏感性。因此，需要改进的诊断工具来支持 STH 控制程序的决策。方法学 我们开发了一种基于重组酶聚合酶扩增 (RPA) 技术的核酸扩增测试，以检测粪便中的 STH。我们为四种 STH 物种中的每一种设计了引物和探针，优化了测定，然后使用临床粪便样本验证其性能。主要发现 在检测克隆的目标 DNA 序列的拷贝时，每个 RPA 检测都与实时聚合酶链反应 (PCR) 检测一样灵敏。基于 Kato-Katz 方法，RPA 分析扩增了从人类粪便样本中提取的 DNA 中的目标，这些样本对 STH 呈阳性，非目标基因组 DNA 没有交叉反应。当对来自轻度、中度和重度感染患者的临床粪便样本进行测试时，RPA 分析显示出与实时 PCR 相当的性能，结果优于 Kato-Katz。这种用于检测 STH 感染的快速、灵敏且可现场部署的新方法可以帮助 STH 控制计划实现其目标。结论 通过 RPA 分析对目标进行半定量是可能的，并且与实时 PCR 相当。借助适当的仪器，RPA 检测可以对 STH DNA 目标进行可靠的半定量，作为评估感染强度的蠕虫卵计数的替代性现场部署指标。