Whole genome amplification and exome sequencing of archived schistosome miracidia

Le Clec'H, W; Chevalier, F D; McDew-White, M; Allan, F; Webster, B L; Gouvras, A N; Kinunghi, S; Tchuem Tchuenté, L-A; Garba, A; Mohammed, K A; Ame, S M; Webster, J P; Rollinson, D; Emery, A M; Anderson, T J

Abstract

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.

Citation

Le Clec'H, W., Chevalier, F. D., McDew-White, M., Allan, F., Webster, B. L., Gouvras, A. N., Kinunghi, S., Tchuem Tchuenté, L.-A., Garba, A., Mohammed, K. A., Ame, S. M., Webster, J. P., Rollinson, D., Emery, A. M., & Anderson, T. J. (2018). Whole genome amplification and exome sequencing of archived schistosome miracidia. Parasitology, 145, 1739-1747. https://doi.org/10.1017/S0031182018000811

• Journal Article Type: Article
• Acceptance Date: Apr 5, 2018
• Publication Date: May 28, 2018
• Deposit Date: Nov 9, 2018
• Publicly Available Date: Nov 9, 2018
• Journal: Parasitology
• Print ISSN: 0031-1820
• Electronic ISSN: 1469-8161
• Publisher: Cambridge University Press
• Peer Reviewed: Peer Reviewed
• Volume: 145
• Pages: 1739-1747
• DOI: https://doi.org/10.1017/S0031182018000811
• Public URL: https://rvc-repository.worktribe.com/output/1387077