Human strongyloidiasis a soil-transmitted infection caused by Strongyloides stercoralis is one of the most neglected amongst the so-called Neglected Tropical Diseases (NTDs). S. stercoralis is a nematode, which is distributed worldwide; it has been estimated that it could affect millions of people, mainly in tropical and subtropical endemic regions. The difficulties of diagnosis lead to infection rates being underreported. Asymptomatic patients have chronic infections that can lead to severe hyperinfection syndrome or disseminated strongyloidiasis in immunocompromised patients. Strongyloidiasis can easily be misdiagnosed because conventional faecal-based techniques lack of sensitivity for the morphological identification of infective larvae in faeces. None of the currently used molecular methods have used urine samples as an alternative to faecal samples for diagnosing strongyloidiasis. This study was thus aimed at comparing, for the first time, the use of a new loop-mediated isothermal amplification (LAMP) molecular assay (Strong-LAMP) to traditional methods on patients’ urine samples. Twenty-four urine samples were taken from patients included in a study involving two Spanish hospitals for strongyloidiasis screening using parasitological and serological tests. Strongyloides larvae were found in 11 patients’ faecal samples, thereby ascertaining that they had the disease. Other patients had high antibody titres but no larvae were found in their faeces. All urine samples were analysed by PCR and Strong-LAMP assay. No amplification occurred when using PCR. Strong-LAMP led to detecting S. stercoralis DNA in urine samples from patients having previously confirmed strongyloidiasis by parasitological tests and/or a suspicion of being infected by serological ones. The Strong-LAMP assay is a useful molecular tool for research regarding strongyloidiasis in human urine samples. After further validation, the Strong-LAMP assay could also be used for complementary and effective diagnosis of strongyloidiasis in a clinical setting.

中文翻译：

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基于18S rRNA中的强线虫属物种衍生的部分序列的Strong-LAMP分析，作为人类尿液样品中强线虫病诊断的潜在生物标记。

人类圆线虫病是一种由土壤固线虫（Strongyloides stercoralis）引起的由土壤传播的感染，是所谓的被忽视热带病（NTDs）中最被忽视的疾病之一。固醇链球菌是一种线虫，分布于世界各地；据估计，它可能影响数百万人，主要是在热带和亚热带流行地区。诊断的困难导致感染率被低估。无症状患者患有慢性感染，可导致免疫功能低下的患者出现严重的过度感染综合症或弥散性线虫病。由于传统的基于粪便的技术对粪便中感染性幼虫的形态学鉴定缺乏敏感性，因此很容易误诊为圆线虫病。当前使用的分子方法均未使用尿液样品作为粪便样品的替代品来诊断强直性根病。因此，本研究旨在首次比较使用新的环介导的等温扩增（LAMP）分子分析（常规方法对患者尿液样本进行检测。从包括在两家西班牙医院进行的一项研究中的患者中抽取了二十四份尿液样本，用于使用寄生虫学和血清学检查筛查弓形虫病。在11名患者的粪便样本中发现了类圆线虫幼虫，从而确定他们患有这种疾病。其他患者的抗体滴度很高，但粪便中未发现幼虫。通过PCR和Strong- LAMP测定法分析所有尿液样品。使用PCR时未发生扩增。Strong- LAMP导致检测粪类圆线虫先前通过寄生虫学检查和/或怀疑被血清学感染确诊为类圆线虫病的患者尿液样本中的DNA。该强-LAMP检测是关于人的尿样中线虫研究的一个有用的分子工具。经过进一步验证后，Strong -LAMP分析还可以在临床环境中用于互补和有效诊断圆线虫病。