Molecular tools to monitor eradication of Schistosoma haematobium transmission

描述（由申请人提供）：目前的大规模血吸虫病治疗计划是减少全球血吸虫病相关疾病负担的第一步，但这些计划可能不会显著改变高危地区的寄生虫传播。因此，这些计划的初始效益的可持续性仍然值得怀疑，因为反复发生的低水平再感染可能与贫血、营养不良和表现下降等持续发病率有关。需要用于快速、灵敏的传播监测的新型分子工具，以获得更多信息的监测扩展区域内的寄生虫传播。雅阁R21计划的发展重点，该项目将创建和验证新技术，这些技术可以显着提高对血吸虫病传播环境特征的认识，并将为改善血吸虫病控制的健康相关实施研究做出重大贡献。该研究的总体目标是测试新S.血吸虫DNA检测分子工具，以有效地监测血吸虫病传播的多社区运动。这种寄生虫在非洲和中东广泛流行，目前有1.2亿人感染。有效的“xenomonitoring”蜗牛采样策略可以提供高度敏感的系统，监测传播潜力的寄生虫控制运动的实施。本研究的环介导等温扩增是一种可扩展的方法，可用于提供低技术含量的DNA检测系统，区域和国际控制计划将能够用作记录其有效性并确认实现血吸虫病根除的手段。本研究的具体目的是：1.为验证已建立的525 bp Sh 110/SmS 1 PCR技术作为一种种特异性的方法来鉴定感染S.在生态系统中，相关的寄生虫物种是同域的。2.目的建立一种低技术含量的钉螺潜隐感染的检测方法，即采用环介导等温扩增（LAMP）技术扩增血吸虫Sh 110/SmSL重复序列。埃及血吸虫3.通过对实验室感染钉螺和现场采集的钉螺进行检测，验证LAMP方法的有效性。肯尼亚的血吸虫病流行区。4.在肯尼亚的一个试点现场实验室和一个中心参考实验室建立LAMP作为工作检测，以进行备份和质量控制。5.评价S.在肯尼亚现有的控制设置中的haematobiom-LAMP性能。人类血吸虫病是一种寄生吸虫感染，对发展中国家造成重大健康负担，需要新的战略来跟踪其通过环境的不均匀传播。这项建议开发了新的DNA检测系统，以监测受感染的中间宿主蜗牛之间的寄生虫传播（通过当地受感染的水体）。这一工具将为指导血吸虫病控制项目提供一种非常有用的方法，并可能导致更有效的根除寄生虫传播的有针对性的策略。

DESCRIPTION (provided by applicant): Current large-scale schistosomiasis treatment programs are a first step to reducing the global burden of Schistosoma-related disease, yet such programs may not significantly alter parasite transmission in high-risk areas. Consequently, the sustainability of these programs' initial benefits remains in doubt, as recurring low-level reinfection can be associated with persistent morbidity such as anemia, undernutrition, and diminished performance. Novel molecular tools for rapid, sensitive transmission monitoring are needed to obtain more informative surveillance of schistosome propagation over extended areas. In accord with the R21 program's developmental focus, this project will create and validate new technologies that can significantly advance knowledge of the environmental features of schistosome transmission, and will materially contribute to health-related implementation research for improved schistosomiasis control. The overall aim of the proposed study is to test the capacity of new S. haematobium DNA-detection molecular tools to efficiently monitor schistosomiasis transmission in multi-community campaigns. The parasite is widely prevalent in Africa and the Middle East, with 120 million persons currently infected. Effective 'xenomonitoring' snail sampling strategies can provide highly sensitive systems for monitoring transmission potential following implementation of parasite control campaigns. The present study's Loop-mediated Isothermal Amplification is a scalable approach can be adapted to provide low-tech DNA detection systems that regional and international control programs will be able use as a means to document their effectiveness and confirm attainment of schistosomiasis eradication. The Specific Aims of the study are: 1. To validate the established 525 bp Sh110/SmSl PCR technique as a species-specific means to identify snails infected with S. haematobium in ecosystems where related schistosome species are sympatric. 2. To develop a low-tech detection of prepatent snail infection by adapting the Loop-Mediated Isothermal Amplification (LAMP) assay for amplifying the inter-repeat 525 bp Sh110/SmSL sequence of S. haematobium. 3. To validate the LAMP assay by examining laboratory infected snails and field snails collected from S. haematobium-endemic areas in Kenya. 4. To establish the LAMP as a working assay in a pilot field laboratory in Kenya and in a central reference laboratory for backup and quality control. 5. To evaluate S. haematobium-LAMP performance in an existing control setting in Kenya. Human schistosomiasis is a parasitic trematode infection that imposes a major health burden on the developing world and new strategies are needed for tracking its uneven transmission through the environment. This proposal develops new DNA detection systems to monitor parasite transmission (through local infested water bodies) among infected intermediate host snails. This tool will provide a highly useful method of directing schistosomiasis control programs, and could lead to much more effective targeted strategies for eradication of parasite transmission.