A microarray platform and novel SNP calling algorithm to evaluate Plasmodium falciparum field samples of low DNA quantity.

Journal Article

Analysis of single nucleotide polymorphisms (SNPs) derived from whole-genome studies allows for rapid evaluation of genome-wide diversity, and genomic epidemiology studies of Plasmodium falciparum provide insights into parasite population structure, gene flow, drug resistance and vaccine development. In areas with adequate cold chain facilities, large volumes of leukocyte-depleted patient blood can be frozen for use in parasite genomic analyses. In more remote endemic areas smaller volumes of infected blood are taken by finger prick, and dried and stored on filter paper. These dried blood spots do not generally yield enough concentrated parasite DNA for whole-genome sequencing.A DNA microarray was designed for use on field samples to type a genome-wide set of SNPs which prior sequencing had shown to be variable in Africa, Southeast Asia, and Papua New Guinea. An algorithm was designed to call SNPs in samples with low parasite DNA. With this new algorithm SNP-calling accuracy of 98% was measured by hybridizing purified DNA from malaria lab strains and comparing calls with SNPs called from full genome sequences. An average accuracy of >98% was likewise obtained for DNA extracted from malaria field samples collected in studies in Southeast Asia, with an average call rate of > 82%.This new high-density microarray provided high quality SNP calls from a wide range of parasite DNA quantities, and represents a robust tool for genome-wide analysis of malaria parasites in diverse settings.

Full Text

Duke Authors

Cited Authors

  • Jacob, CG; Tan, JC; Miller, BA; Tan, A; Takala-Harrison, S; Ferdig, MT; Plowe, CV

Published Date

  • August 26, 2014

Published In

Volume / Issue

  • 15 /

Start / End Page

  • 719 -

PubMed ID

  • 25159520

Electronic International Standard Serial Number (EISSN)

  • 1471-2164

International Standard Serial Number (ISSN)

  • 1471-2164

Digital Object Identifier (DOI)

  • 10.1186/1471-2164-15-719

Language

  • eng