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Single-cell genomics is a powerful tool for determining the genetic architecture of complex communities of unicellular organisms. In areas of high transmission, malaria patients are often challenged by the activities of multiple Plasmodium falciparum lineages, which can potentiate pathology, spread drug resistance loci, and also complicate most genetic analysis. Single-cell sequencing of P. falciparum would be key to understanding infection complexity, though efforts are hampered by the extreme nucleotide composition of its genome (∼80% AT-rich). To counter the low coverage achieved in previous studies, we targeted DNA-rich late-stage parasites by Fluorescence-Activated Cell Sorting and whole genome sequencing. Our method routinely generates accurate, near-complete capture of the 23 Mb P. falciparum genome (mean breadth of coverage 90.7%) at high efficiency. Data from 48 single-cell genomes derived from a polyclonal infection sampled in Chikhwawa, Malawi allowed for unambiguous determination of haplotype diversity and recent meiotic events, information that will aid public health efforts.

Original publication

DOI

10.1093/gbe/evx256

Type

Journal article

Journal

Genome Biol Evol

Publication Date

01/12/2017

Volume

9

Pages

3373 - 3383

Keywords

malaria, methods, single-cell genomics, Child, Preschool, DNA, Protozoan, Erythrocytes, Genetic Variation, Genome, Protozoan, Haplotypes, Humans, Malaria, Falciparum, Malawi, Plasmodium falciparum, Polymerase Chain Reaction, Single-Cell Analysis