Malaga Jose L, Fernandez-Baca Martha V, Castellanos-Gonzalez Alejandro, Tanabe Melinda B, Tift Clara, Morales Maria Luisa, Lopez Martha, Valdivia-Rodriguez Angela, Mamani-Licona Frecia, Cabada Miguel M
Sede Cusco-Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima 15102, Peru.
Universidad Peruana Cayetano Heredia-University of Texas Medical Branch Collaborative Research Center-Cusco, Universidad Peruana Cayetano Heredia, Cusco 08002, Peru.
Pathogens. 2024 Oct 3;13(10):869. doi: 10.3390/pathogens13100869.
Strongyloidiasis is a neglected, soil-transmitted helminth infection prevalent worldwide. The true burden of strongyloidiasis is unclear due to the lack of sensitive, field-friendly diagnostic tests. PCR tests to detect DNA in stool are sensitive and specific, but the need for expensive equipment limits their use in endemic regions. Isothermal PCR amplification tests are easier to deploy while maintaining sensitivity and specificity. We developed and evaluated a recombinase polymerase amplification lateral flow assay (RPA-LFA) to detect in human stool samples. Three hundred stool samples were collected in three communities in the jungle of Cusco, Peru. Samples were tested for larvae using microscopy (Baermann's, agar plate culture (APC), and rapid sedimentation), real-time PCR, and RPA-LF for DNA. The RPA-LFA showed an analytical limit of detection of 20 pg/µL. The prevalence of was 27%, 38%, 46.3%, and 46% using microscopy, PCR, microscopy/PCR, and RPA-LFA, respectively. RPA-LFA had a sensitivity and specificity of 59.3% and 58.9%, 66.2% and 71.4%, and 77.2% and 73.1% when microscopy, microscopy/PCR, and real-time PCR were used as the gold standards, respectively. The RPA-LFA is a novel, fast, highly sensitive, and specific molecular method with the potential for deployment in endemic regions.
粪类圆线虫病是一种被忽视的、经土壤传播的蠕虫感染，在全球范围内普遍存在。由于缺乏灵敏且便于现场使用的诊断测试，粪类圆线虫病的真实负担尚不清楚。用于检测粪便中DNA的PCR测试灵敏且特异，但因其需要昂贵的设备，限制了其在流行地区的使用。等温PCR扩增测试在保持灵敏性和特异性的同时更易于部署。我们开发并评估了一种重组酶聚合酶扩增侧向流动分析法（RPA-LFA），用于检测人类粪便样本中的粪类圆线虫。在秘鲁库斯科丛林的三个社区收集了300份粪便样本。使用显微镜检查（贝尔曼法、琼脂平板培养法（APC）和快速沉淀法）、实时PCR以及用于检测粪类圆线虫DNA的RPA-LF对样本进行粪类圆线虫幼虫检测。RPA-LFA的分析检测限为20 pg/µL。使用显微镜检查、PCR、显微镜检查/PCR和RPA-LFA检测粪类圆线虫的患病率分别为27%、38%、46.3%和46%。当分别以显微镜检查、显微镜检查/PCR和实时PCR作为金标准时，RPA-LFA的灵敏度和特异性分别为59.3%和58.9%、66.2%和71.4%、77.2%和73.1%。RPA-LFA是一种新型、快速、高度灵敏且特异的分子方法，有在流行地区部署的潜力。
