Efficient genotyping of Schistosoma mansoni miracidia following whole genome amplification

Claudia L.L. Valentim, Philip T. LoVerde, Timothy JC Anderson, Charles D. Criscione

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)81-84
Number of pages4
JournalMolecular and Biochemical Parasitology
Volume166
Issue number1
DOIs
StatePublished - Jul 2009

Keywords

  • Mendelian
  • Microsatellites
  • Miracidia
  • Multiple displacement amplification
  • Schistosoma

ASJC Scopus subject areas

  • Parasitology
  • Molecular Biology

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