Whole genome amplification and exome sequencing of archived schistosome miracidia

Winka Le Clec'h, Frédéric D. Chevalier, Marina McDew-White, Fiona Allan, Bonnie L. Webster, Anouk N. Gouvras, Safari Kinunghi, Louis Albert Tchuem Tchuenté, Amadou Garba, Khalfan A. Mohammed, Shaali M. Ame, Joanne P. Webster, David Rollinson, Aidan M. Emery, Timothy J.C. Anderson

    Research output: Contribution to journalArticlepeer-review

    8 Scopus citations

    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.

    Original languageEnglish (US)
    Pages (from-to)1739-1747
    Number of pages9
    JournalParasitology
    Volume145
    Issue number13
    DOIs
    StatePublished - Nov 1 2018

    Keywords

    • Exome sequencing
    • FTA cards
    • Schistosoma
    • miracidia
    • quantitative PCR
    • whole genome amplification

    ASJC Scopus subject areas

    • Parasitology
    • Animal Science and Zoology
    • Infectious Diseases

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