Efficient genotyping of Schistosoma mansoni miracidia following whole genome amplification

Small parasites and larval stages pose a problem for molecular analyses because limited amounts of DNA template are available. Isothermal methods for faithfully copying DNA have the potential to revolutionize studies of such organisms. We evaluated the fidelity of multiple displacement amplification...

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Bibliographic Details
Published in:Molecular and biochemical parasitology 2009-07, Vol.166 (1), p.81-84
Main Authors: Valentim, Claudia L.L., LoVerde, Philip T., Anderson, Timothy J.C., Criscione, Charles D.
Format: Article
Language:English
Subjects:
Animals
Female
Genetic Techniques - standards
Genome, Helminth - genetics
Genotype
Life Cycle Stages
Male
Mendelian
Microsatellite Repeats - genetics
Microsatellites
Miracidia
Multiple displacement amplification
Reproducibility of Results
Schistosoma
Schistosoma mansoni
Schistosoma mansoni - genetics
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Summary:Small parasites and larval stages pose a problem for molecular analyses because limited amounts of DNA template are available. Isothermal methods for faithfully copying DNA have the potential to revolutionize studies of such organisms. We evaluated the fidelity of multiple displacement amplification (MDA) for amplifying DNA extracted from a single miracidium of Schistosoma mansoni. To do this we genotyped DNA extracted from 28 F1 miracidia following MDA using 56 microsatellite markers. Because these miracidia were obtained from a cross between a male and female worm of known genotypes, we were able to predict the alleles present in the progeny and quantify the genotyping error rate. We found just 8/1568 genotypes deviated from Mendelian expectations. Furthermore, because 1 of these resulted from a genuine mutation, the error rate due to MDA is 7/1568 (0.45%). We conclude that many hundreds of microsatellites or other genetic markers can be accurately genotyped from a single miracidium using this method, greatly expanding the scope of population genetic, epidemiological and evolutionary studies on this parasite.
ISSN:0166-6851
1872-9428
DOI:10.1016/j.molbiopara.2009.02.010