Plasmodium vivax: microsatellite analysis of multiple-clone infections |
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Authors: | Havryliuk Tatiana Orjuela-Sánchez Pamela Ferreira Marcelo U |
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Institution: | aDepartment of Parasitology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil;bMount Sinai School of Medicine, New York, NY, USA |
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Abstract: | 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. |
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Keywords: | Plasmodium vivax Multiple-clone infection Malaria Genetic diversity Microsatellites bp base pairs DNA deoxyribonucleic acid FU fluorescence units MS microsatellite MSP1 merozoite surface protein 1 PCR polymerase chain reaction WGA whole-genome amplification |
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