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

Author:
  • Javier Perez-Tris
  • Staffan Bensch
Publishing year: 2005
Language: English
Pages: 15-23
Publication/Series: Parasitology
Volume: 131
Issue: 1
Document type: Journal article
Publisher: Cambridge University Press

Abstract english

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.

Keywords

  • Biological Sciences

Other

Published
  • Molecular Ecology and Evolution Lab
  • ISSN: 1469-8161
Staffan Bensch
E-mail: staffan [dot] bensch [at] biol [dot] lu [dot] se

Professor

MEMEG

+46 46 222 42 92

E-C213

Sölvegatan 37, Lund

50

Head of unit

MEMEG

+46 46 222 42 92

E-C213

Sölvegatan 37, Lund

50

Research group

Molecular Ecology and Evolution Lab

Projects

Doctoral students and postdocs

PhD students, main supervisor

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