Plasmodium vivax: microsatellite analysis of multiple-clone infections.
We used mixtures of genomic DNA from two genetically distinct isolates from Brazil, 42M and 312M, to investigate how accurately 12-locus microsatellite typing describes the overall genetic diversity and characterizes multilocus haplotypes in multiple-clone Plasmodium vivax infections. We found varying PCR amplification efficiencies of microsatellite alleles; for example, from the same 1:1 mixture of 42M and 312M DNA we amplified predominantly 312M-type alleles at 10 loci and 42M-type alleles at 2 loci. All microsatellite alleles were accurately scored in 1:0.5 and 1:0.25 312M:42M DNA mixtures, even when minor peak heights did not meet previously suggested criteria for minor allele detection in multiple-clone infections. Relative proportions of major and minor alleles were unaffected by multiple displacement amplification of template DNA prior to PCR-based microsatellite typing. Although microsatellite typing may detect minor alleles in clone mixtures, amplification biases may lead to inaccurate assignment of predominant haplotypes in multiple-clone P. vivax infections.
Havryliuk
Tatiana
T
Department of Parasitology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
Orjuela-Sánchez
Pamela
P
Ferreira
Marcelo U
MU
D000483:Alleles; D000818:Animals; D016054:DNA, Protozoan / Q000032:analysis*; D005784:Gene Amplification; D018503:Genome, Protozoan / Q000235:genetics; D006239:Haplotypes / Q000235:genetics; D006801:Humans; D016780:Malaria, Vivax / Q000469:parasitology*; D018895:Microsatellite Repeats / Q000235:genetics*; D010966:Plasmodium vivax / Q000145:classification* / Q000235:genetics*; D016133:Polymerase Chain Reaction
D016054:DNA, Protozoan