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Nested cytochrome b polymerase chain reaction diagnostics underestimate mixed infections of avian blood haemosporidian parasites: microscopy is still essential.

Abstract
Numerous polymerase chain reaction (PCR)-based methods have been developed and used increasingly to screen vertebrate blood samples for the diagnosis of haemosporidian blood parasites (Sporozoa, Haemosporida), but a rigorous evaluation of the sensitivity of these methods for detecting mixed infections of different haemosporidian species belonging to the same and different genera and subgenera is lacking. This study links the information obtained by nested cytochrome b PCR and traditional microscopy in determining mixed haemosporidian infections in naturally infected birds. Samples from 83 individual passerine birds with single infections of Haemoproteus or Plasmodium spp., as determined by mitochondrial DNA amplification, also were investigated by microscopic examination of stained blood films. Thirty-six samples (43%) were found to harbor mixed Haemoproteus, or Plasmodium spp. infections, or both. Thus, the PCR assays alone underestimate the occurrence of mixed infections of haemosporidian parasites in naturally infected birds. To determine the true species composition of the haemosporidians in each individual host, PCR diagnostics need to be improved. Specific primers for Haemoproteus spp. and Plasmodium spp. should be developed. Ideally, a combination of the approaches of both microscopy and PCR-based methods is recommended for this purpose.
AuthorsGediminas Valkiŭnas, Staffan Bensch, Tatjana A Iezhova, Asta Krizanauskiené, Olof Hellgren, Casimir V Bolshakov
JournalThe Journal of parasitology (J Parasitol) Vol. 92 Issue 2 Pg. 418-22 (Apr 2006) ISSN: 0022-3395 [Print] United States
PMID16729711 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)
Chemical References
  • DNA, Mitochondrial
  • DNA, Protozoan
  • Cytochromes b
Topics
  • Animals
  • Bird Diseases (diagnosis, parasitology)
  • Cytochromes b (genetics)
  • DNA, Mitochondrial (analysis)
  • DNA, Protozoan (analysis)
  • Haemosporida (enzymology, genetics, isolation & purification)
  • Passeriformes (parasitology)
  • Polymerase Chain Reaction (standards, veterinary)
  • Protozoan Infections, Animal (diagnosis, parasitology)
  • Sensitivity and Specificity
  • Species Specificity

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