Molecular Fingerprinting of Schistosoma Miracidia

Molecular analysis was carried out on up to 20 miracidia per child from the 2010 collections depending on the infection intensity of the child. We replicated the identical methodologies of the original 2005 and 2006 surveys in order that the data were directly comparable [13 (link)], since the original miracidial samples were no longer available for re-analysis. In brief, DNA preparation was carried out on the Whatman cards as per the manufacturers protocol (Whatman FTA cards®). PCR was carried out using the previously published multiplex assay using seven pairs of microsatellite primers [15 (link)], namely SMD28, SMDA28, SDM25, SMD89, CA11-1, SMS9-1 and SMU31768. Forward primers were labelled using 6-FAM, PET, VIC and NED dyes. Microsatellite sizing was performed on the same ABI Prism 3730 Genetic Analyser with a LIZ-500 size standard (Applied Biosystems, Cheshire, UK) as the 2005 and 2006 samples.