Le Clec'h Winka, Chevalier Frédéric D, McDew-White Marina, Allan Fiona, Webster Bonnie L, Gouvras Anouk N, Kinunghi Safari, Tchuem Tchuenté Louis-Albert, Garba Amadou, Mohammed Khalfan A, Ame Shaali M, Webster Joanne P, Rollinson David, Emery Aidan M, Anderson Timothy J C
Department of Genetics,Texas Biomedical Research Institute,PO Box 760549, San Antonio, TX 78245-0549,USA.
Department of Life Sciences,The Natural History Museum,Cromwell Road, London SW7 5BD,UK.
Parasitology. 2018 Nov;145(13):1739-1747. doi: 10.1017/S0031182018000811. Epub 2018 May 28.
Adult schistosomes live in the blood vessels and cannot easily be sampled from humans, so archived miracidia larvae hatched from eggs expelled in feces or urine are commonly used for population genetic studies. Large collections of archived miracidia on FTA cards are now available through the Schistosomiasis Collection at the Natural History Museum (SCAN). Here we describe protocols for whole genome amplification of Schistosoma mansoni and Schistosome haematobium miracidia from these cards, as well as real time PCR quantification of amplified schistosome DNA. We used microgram quantities of DNA obtained for exome capture and sequencing of single miracidia, generating dense polymorphism data across the exome. These methods will facilitate the transition from population genetics, using limited numbers of markers to population genomics using genome-wide marker information, maximising the value of collections such as SCAN.
成年血吸虫生活在血管中，不易从人体采集样本，因此，通常使用从粪便或尿液中排出的虫卵孵化出的保存毛蚴幼虫进行群体遗传学研究。通过自然历史博物馆的血吸虫病藏品库（SCAN），现在可以获得大量保存在FTA卡上的毛蚴。在这里，我们描述了从这些卡片上对曼氏血吸虫和埃及血吸虫毛蚴进行全基因组扩增的方案，以及对扩增的血吸虫DNA进行实时PCR定量的方法。我们使用微克级的DNA进行单个毛蚴的外显子捕获和测序，在整个外显子上生成密集的多态性数据。这些方法将有助于从使用有限数量标记的群体遗传学向使用全基因组标记信息的群体基因组学转变，从而最大限度地提高SCAN等藏品库的价值。
