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Parasitol Res. 2016 Dec;115(12):4503-4510. Epub 2016 Sep 21.

Laser capture microdissection microscopy and genome sequencing of the avian malaria parasite, Plasmodium relictum.

Author information

1
Department of Ecology and Evolutionary Biology, College of Agricultural and Life Sciences, Cornell University, Ithaca, NY, 14853, USA. hlutz@fieldmuseum.org.
2
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, 14853, USA. hlutz@fieldmuseum.org.
3
Cornell Lab of Ornithology, Cornell University, Ithaca, NY, 14853, USA. hlutz@fieldmuseum.org.
4
Integrative Research Center, The Field Museum of Natural History, Chicago, IL, 60605, USA. hlutz@fieldmuseum.org.
5
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, 14853, USA.
6
Integrative Research Center, The Field Museum of Natural History, Chicago, IL, 60605, USA.
7
Department of Entomology, University of California, Davis, CA, 95616, USA.
8
Nature Research Centre, Akademijos 2, Vilnius, LT-08412, Lithuania.
9
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, 14853, USA. mjs297@cornell.edu.

Abstract

Acquiring genomic material from avian malaria parasites for genome sequencing has proven problematic due to the nucleation of avian erythrocytes, which produces a large ratio of host to parasite DNA (∼1 million to 1 bp). We tested the ability of laser capture microdissection microscopy to isolate parasite cells from individual avian erythrocytes for four avian Plasmodium species, and subsequently applied whole genome amplification and Illumina sequencing methods to Plasmodium relictum (lineage pSGS1) to produce sequence reads of the P. relictum genome. We assembled ∼335 kbp of parasite DNA from this species, but were unable to completely avoid contamination by host DNA and other sources. However, it is clear that laser capture microdissection holds promise for the isolation of genomic material from haemosporidian parasites in intracellular life stages. In particular, laser capture microdissection may prove useful for isolating individual parasite species from co-infected hosts. Although not explicitly tested in this study, laser capture microdissection may also have important applications for isolation of rare parasite lineages and museum specimens for which no fresh material exists.

KEYWORDS:

Avian malaria; Plasmodium genome; laser capture microdissection microscopy

PMID:
27651044
DOI:
10.1007/s00436-016-5237-5
[Indexed for MEDLINE]

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