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A restriction site to differentiate Plasmodium and Haemoproteus infections in birds: on the inefficiency of general primers for detection of mixed infections

Published online by Cambridge University Press:  06 May 2009

J. MARTÍNEZ ,
J. MARTÍNEZ-DE LA PUENTE ,
J. HERRERO ,
S. DEL CERRO ,
E. LOBATO ,
J. RIVERO-DE AGUILAR ,
R. A. VÁSQUEZ  and
S. MERINO
J. MARTÍNEZ*
Affiliation:
Departamento de Microbiología y Parasitología, Facultad de Farmacia, Universidad de Alcalá, Alcalá de Henares, E-28871 Madrid, Spain
J. MARTÍNEZ-DE LA PUENTE
Affiliation:
Departamento de Ecología Evolutiva, Museo Nacional de Ciencias Naturales-CSIC, J. Gutiérrez Abascal 2, E-28006 Madrid, Spain
J. HERRERO
Affiliation:
Departamento de Microbiología y Parasitología, Facultad de Farmacia, Universidad de Alcalá, Alcalá de Henares, E-28871 Madrid, Spain
S. DEL CERRO
Affiliation:
Departamento de Ecología Evolutiva, Museo Nacional de Ciencias Naturales-CSIC, J. Gutiérrez Abascal 2, E-28006 Madrid, Spain
E. LOBATO
Affiliation:
Departamento de Ecología Evolutiva, Museo Nacional de Ciencias Naturales-CSIC, J. Gutiérrez Abascal 2, E-28006 Madrid, Spain
J. RIVERO-DE AGUILAR
Affiliation:
Departamento de Ecología Evolutiva, Museo Nacional de Ciencias Naturales-CSIC, J. Gutiérrez Abascal 2, E-28006 Madrid, Spain
R. A. VÁSQUEZ
Affiliation:
Instituto de Ecología y Biodiversidad, Departamento de Ciencias Ecológicas, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Ñuñoa, Santiago, Chile
S. MERINO
Affiliation:
Departamento de Ecología Evolutiva, Museo Nacional de Ciencias Naturales-CSIC, J. Gutiérrez Abascal 2, E-28006 Madrid, Spain
*
*Corresponding author: Departamento de Microbiología y Parasitología, Facultad de Farmacia, Universidad de Alcalá, Alcalá de Henares, E-28871 Madrid, Spain. Tel: +34 918854636. Fax: +34 918854663. E-mail: francisco.martinez@uah.es
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Summary

Avian Plasmodium and Haemoproteus parasites are easily detected by DNA analyses of infected samples but only correctly assigned to each genus by sequencing and use of a phylogenetic approach. Here, we present a restriction site to differentiate between both parasite genera avoiding the use of those analyses. Alignments of 820 sequences currently listed in GenBank encoding a particular cytochrome B region of avian Plasmodium and Haemoproteus show a shared restriction site for both genera using the endonuclease Hpy CH4III. An additional restriction site is present in Plasmodium sequences that would initially allow differentiation of both genera by differential migration of digested products on gels. Overall 9 out of 326 sequences containing both potential restriction sites do not fit to the general rule. We used this differentiation of parasite genera based on Hpy CH4III restriction sites to evaluate the efficacy of 2 sets of general primers in detecting mixed infections. To do so, we used samples from hosts infected by parasites of both genera. The use of general primers was only able to detect 25% or less of the mixed infections. Therefore, parasite DNA amplification using general primers to determine the species composition of haemosporidian infections in individual hosts is not recommended. Specific primers for each species and study area should be designed until a new method can efficiently discriminate both parasites.

Keywords

avian malariaendonuclease Hpy CH4IIIhaemoparasitesHaemosporidiahost-parasite interactionsmixed infectionsmolecular detectionPCR
Type
Research Article
Information
Parasitology , Volume 136 , Issue 7 , June 2009 , pp. 713 - 722
Copyright
Copyright © Cambridge University Press 2009

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