Advanced

Label-free enrichment of avian Leucocytozoon using flow cytometric sorting

Chakarov, Nayden LU ; Greiner, J. F. W.; Hauser, S.; Schuetzmann, D.; Krüger, O.; Boerner, M. and Widera, D. (2012) In Parasitology 139(12). p.1547-1552
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... (More)
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. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Leucocytozoon, flow cytometry, label-free enrichment, plasmodium-falciparum, malaria, identification, gametocytes, separation, blood
in
Parasitology
volume
139
issue
12
pages
1547 - 1552
publisher
Cambridge University Press
external identifiers
  • scopus:84867799996
ISSN
1469-8161
DOI
10.1017/s0031182012000972
language
English
LU publication?
no
id
a1610f6a-04a8-4917-894c-3f6b9dfc2b1e (old id 4936924)
date added to LUP
2015-01-22 11:19:15
date last changed
2017-01-01 07:35:32
@article{a1610f6a-04a8-4917-894c-3f6b9dfc2b1e,
  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.},
  author       = {Chakarov, Nayden and Greiner, J. F. W. and Hauser, S. and Schuetzmann, D. and Krüger, O. and Boerner, M. and Widera, D.},
  issn         = {1469-8161},
  keyword      = {Leucocytozoon,flow cytometry,label-free enrichment,plasmodium-falciparum,malaria,identification,gametocytes,separation,blood},
  language     = {eng},
  number       = {12},
  pages        = {1547--1552},
  publisher    = {Cambridge University Press},
  series       = {Parasitology},
  title        = {Label-free enrichment of avian Leucocytozoon using flow cytometric sorting},
  url          = {http://dx.doi.org/10.1017/s0031182012000972},
  volume       = {139},
  year         = {2012},
}