Label-free enrichment of avian Leucocytozoon using flow cytometric sorting
(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:
https://lup.lub.lu.se/record/4936924
- author
- Chakarov, Nayden LU ; Greiner, J. F. W. ; Hauser, S. ; Schuetzmann, D. ; Krüger, O. ; Boerner, M. and Widera, D.
- organization
- publishing date
- 2012
- 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
- 2016-04-04 08:14:28
- date last changed
- 2022-01-29 03:10:38
@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}},
keywords = {{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}},
doi = {{10.1017/s0031182012000972}},
volume = {{139}},
year = {{2012}},
}