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Diagnosing genetically diverse avian malarial infections using mixed-sequence analysis and TA-cloning

Perez-Tris, Javier LU and Bensch, Staffan LU (2005) In Parasitology 131(1). p.15-23
Abstract
Birds harbouring several malarial parasites are common in the wild, and resolving such multiple infections is important for our understanding of host-parasite relationships. We propose a simple and reasonably accurate method for detecting and resolving multiple infections, based on the analysis of parasite cytochrome b DNA sequences: genetically mixed infections are first identified by double nucleotide peaks on sequence electropherograms, and later retrieved by TA-cloning. We applied this method to wild birds, and to experimentally created mixes with varying proportion of two parasites (Plasmodium spp. and Haemoproteus spp.). In general, the method was very efficient in detecting and resolving multiple infections, but some problems were... (More)
Birds harbouring several malarial parasites are common in the wild, and resolving such multiple infections is important for our understanding of host-parasite relationships. We propose a simple and reasonably accurate method for detecting and resolving multiple infections, based on the analysis of parasite cytochrome b DNA sequences: genetically mixed infections are first identified by double nucleotide peaks on sequence electropherograms, and later retrieved by TA-cloning. We applied this method to wild birds, and to experimentally created mixes with varying proportion of two parasites (Plasmodium spp. and Haemoproteus spp.). In general, the method was very efficient in detecting and resolving multiple infections, but some problems were encountered. Several multiple infections were erroneously scored as simple, either because one of the parasite lineages was a better target for the primers used, or because it was much more abundant in the mix. On the other hand, single nucleotide substitutions and template switching during PCR produced artificial sequences in some clones. We discuss the utility of the method, and propose a framework for its use when screening for genetically diverse avian malarial parasites. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Parasitology
volume
131
issue
1
pages
15 - 23
publisher
Cambridge University Press
external identifiers
  • pmid:16038392
  • wos:000231004500002
  • scopus:22444441625
ISSN
1469-8161
DOI
10.1017/S003118200500733X
language
English
LU publication?
yes
id
9a191e6f-856b-4b7b-8ff3-2652a212cd38 (old id 145370)
date added to LUP
2007-06-25 15:32:36
date last changed
2017-10-01 03:36:45
@article{9a191e6f-856b-4b7b-8ff3-2652a212cd38,
  abstract     = {Birds harbouring several malarial parasites are common in the wild, and resolving such multiple infections is important for our understanding of host-parasite relationships. We propose a simple and reasonably accurate method for detecting and resolving multiple infections, based on the analysis of parasite cytochrome b DNA sequences: genetically mixed infections are first identified by double nucleotide peaks on sequence electropherograms, and later retrieved by TA-cloning. We applied this method to wild birds, and to experimentally created mixes with varying proportion of two parasites (Plasmodium spp. and Haemoproteus spp.). In general, the method was very efficient in detecting and resolving multiple infections, but some problems were encountered. Several multiple infections were erroneously scored as simple, either because one of the parasite lineages was a better target for the primers used, or because it was much more abundant in the mix. On the other hand, single nucleotide substitutions and template switching during PCR produced artificial sequences in some clones. We discuss the utility of the method, and propose a framework for its use when screening for genetically diverse avian malarial parasites.},
  author       = {Perez-Tris, Javier and Bensch, Staffan},
  issn         = {1469-8161},
  language     = {eng},
  number       = {1},
  pages        = {15--23},
  publisher    = {Cambridge University Press},
  series       = {Parasitology},
  title        = {Diagnosing genetically diverse avian malarial infections using mixed-sequence analysis and TA-cloning},
  url          = {http://dx.doi.org/10.1017/S003118200500733X},
  volume       = {131},
  year         = {2005},
}