Label-free enrichment of avian Leucocytozoon using flow cytometric sorting.

Abstract:

:The group of haemosporidian parasites is of general interest to basic and applied science, since several species infect mammals, leading to malaria and associated disease symptoms. Although the great majority of haemosporidian parasites appear in bird hosts, as in the case of Leucocytozoon buteonis, there is little genomic information about genetic aspects of their co-evolution with hosts. Consequently, there is a high need for parasite-enrichment strategies enabling further analyses of the genomes, namely without exposure to DNA-intercalating dyes. Here, we used flow cytometry without an additional labelling step to enrich L. buteonis from infected buzzard blood. A specific, defined area of two-dimensional scattergramms was sorted and the fraction was further analysed. The successful enrichment of L. buteonis in the sorted fraction was demonstrated by Giemsa-staining and qPCR revealing a clear increase of parasite-specific genes, while host-specific genes were significantly decreased. This is the first report describing a labelling-free enrichment approach of L. buteonis from infected buzzard blood. The enrichment of parasites presented here is free of nucleic acid-intercalating dyes which may interfere with fluorescence-based methods or subsequent sequencing approaches.

journal_name

Parasitology

journal_title

Parasitology

authors

Chakarov N,Greiner JF,Hauser S,Schuetzmann D,Krüger O,Boerner M,Wıdera D

doi

10.1017/S0031182012000972

subject

Has Abstract

pub_date

2012-10-01 00:00:00

pages

1547-52

issue

12

eissn

0031-1820

issn

1469-8161

pii

S0031182012000972

journal_volume

139

pub_type

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