Webster B L, Rabone M, Pennance T, Emery A M, Allan F, Gouvras A, Knopp S, Garba A, Hamidou A A, Mohammed K A, Ame S M, Rollinson D, Webster J P
Wolfson Wellcome Biomedical Laboratories, Department of Life Sciences, Natural History Museum, Cromwell Road, London, SW7 5BD, UK.
Department of Infectious Disease Epidemiology, Imperial College Faculty of Medicine (St Mary's Campus), Norfolk Place, London, W2 1PG, UK.
Parasit Vectors. 2015 Aug 20;8:432. doi: 10.1186/s13071-015-1044-6.
Human urogenital schistosomiasis caused by Schistosoma haematobium is widely distributed across Africa and is increasingly targeted for control and regional elimination. The development of new high-throughput, cost-effective molecular tools and approaches are needed to monitor and evaluate the impact of control programs on the parasite populations. Microsatellite loci are genetic markers that can be used to investigate how parasite populations change over time and in relation to external influences such as control interventions.
Here, 18 existing S. haematobium microsatellite loci were optimised to enable simultaneous amplification across two novel multiplex microsatellite PCR's, each containing nine loci. Methods were developed for the cost effective and rapid processing and microsatellite analysis of S. haematobium larval stages stored on Whatman-FTA cards and proved robust on miracidia and cercariae collected from Zanzibar and Niger.
The development of these novel and robust multiplex microsatellite assays, in combination with an improved protocol to elute gDNA from Whatman-FTA fixed schistosome larval stages, enables the high-throughput population genetic analysis of S. haematobium. The molecular resources and protocols described here advance the way researchers can perform multi locus-based population genetic analyses of S. haematobium as part of the evaluation and monitoring of schistosomiasis control programmes.
由埃及血吸虫引起的人类泌尿生殖系统血吸虫病广泛分布于非洲，并且越来越多地成为控制和区域消除的目标。需要开发新的高通量、具有成本效益的分子工具和方法来监测和评估控制项目对寄生虫种群的影响。微卫星位点是一种遗传标记，可用于研究寄生虫种群如何随时间变化以及与诸如控制干预等外部影响的关系。
在此，对18个现有的埃及血吸虫微卫星位点进行了优化，以实现跨两个新型多重微卫星PCR同时扩增，每个PCR包含9个位点。开发了用于对保存在Whatman-FTA卡上的埃及血吸虫幼虫阶段进行经济高效且快速处理和微卫星分析的方法，并在从桑给巴尔和尼日尔收集的毛蚴和尾蚴上证明了其可靠性。
这些新型且可靠的多重微卫星检测方法的开发，结合从Whatman-FTA固定的血吸虫幼虫阶段洗脱基因组DNA的改进方案，能够对埃及血吸虫进行高通量群体遗传学分析。本文所述的分子资源和方案推动了研究人员在血吸虫病控制项目的评估和监测中对埃及血吸虫进行基于多位点的群体遗传学分析的方式。
