Detecting number of clones, and their relative abundance, of a malaria parasite (Plasmodium mexicanum) infecting its vertebrate host (original) (raw)
Abstract
Microsatellites, short tandem repeats of nucleotides in the genome, are useful markers to detect clonal diversity within Plasmodium infections. However, accuracy in determining number of clones and their relative proportions based on standard genetic analyzer instruments is poorly known. DNA extracted from lizards infected with a malaria parasite, Plasmodium mexicanum, provided template to genotype the parasite based on three microsatellite markers. Replicate genotyping of the same natural infections demonstrated strong repeatability of data from the instrument. Mixing DNA extracted from several infected lizards simulated mixed-clone infections with known clonal diversity and relative proportions of clones (N = 56 simulations). The instrument readily detected at least four alleles (clones), even when DNA concentrations among clones differed up to tenfold, but alleles of similar size can be missed because they fall within the “stutter” artifact, and rarely does an allele fail to be detected. For simulations of infections that changed their relative proportions over time, changes in relative peak heights on the instrument output closely followed the known changes in relative proportions. Such data are useful for a broad range of studies on the ecology of malaria parasites.
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Acknowledgments
This work was supported by grants from the USA NSF and the Vermont Genetics Network. The Vermont Cancer Center provided valuable technical assistance. We thank B. Calsbeek for important assistance in conducting the experiments and C. W. Kilpatrick for advice throughout the study. All experiments conformed to a protocol from the Animal Care and Use Committee of the University of Vermont and collecting permits from the state of California.
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Authors and Affiliations
- Program in Public Health, University of California at Irvine, Hewitt Hall Room 3501, Irvine, CA, 92697, USA
Anne M. Vardo-Zalik - Department of Biology, University of Vermont, Burlington, VT, 05405, USA
Alice Flynn Ford & Jos. J. Schall
Authors
- Anne M. Vardo-Zalik
- Alice Flynn Ford
- Jos. J. Schall
Corresponding author
Correspondence toJos. J. Schall.
Additional information
Funded by: USA National Science Foundation and the Vermont Genetics Network.
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Vardo-Zalik, A.M., Ford, A.F. & Schall, J.J. Detecting number of clones, and their relative abundance, of a malaria parasite (Plasmodium mexicanum) infecting its vertebrate host.Parasitol Res 105, 209–215 (2009). https://doi.org/10.1007/s00436-009-1385-1
- Received: 10 July 2008
- Accepted: 26 February 2009
- Published: 10 March 2009
- Issue date: June 2009
- DOI: https://doi.org/10.1007/s00436-009-1385-1