A new one-step multiplex PCR assay for simultaneous detection and identification of avian haemosporidian parasites

Research output: Contribution to journalArticle

Abstract

Accurate detection and identification are essential components for epidemiological, ecological, and evolutionary surveys of avian haemosporidian parasites. Microscopy has been used for more than 100 years to detect and identify these parasites; however, this technique requires considerable training and high-level expertise. Several PCR methods with highly sensitive and specific detection capabilities have now been developed in addition to microscopic examination. However, recent studies have shown that these molecular protocols are insufficient at detecting mixed infections of different haemosporidian parasite species and genetic lineages. In this study, we developed a simple, sensitive, and specific multiplex PCR assay for simultaneous detection and discrimination of parasites of the genera Plasmodium, Haemoproteus, and Leucocytozoon in single and mixed infections. Relative quantification of parasite DNA using qPCR showed that the multiplex PCR can amplify parasite DNA ranging in concentration over several orders of magnitude. The detection specificity and sensitivity of this new multiplex PCR assay were also tested in two different laboratories using previously screened natural single and mixed infections. These findings show that the multiplex PCR designed here is highly effective at identifying both single and mixed infections from all three genera of avian haemosporidian parasites. We predict that this one-step multiplex PCR assay, being convenient and inexpensive, will become a widely used method for molecular screening of avian haemosporidian parasites.

Details

Authors
Organisations
External organisations
  • Erciyes University
  • State Scientific Institute Nature Research Centre
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Microbiology

Keywords

  • Haemoproteus, Leucocytozoon, Mixed infection, Multiplex PCR, Parasite detection, Plasmodium
Original languageEnglish
Pages (from-to)191-201
JournalParasitology Research
Volume118
Issue number1
Early online date2018 Dec
Publication statusPublished - 2019 Jan
Publication categoryResearch
Peer-reviewedYes