A New Nested Polymerase Chain Reaction Method Very Efficient in Detecting Plasmodium and Haemoproteus Infections From Avian Blood

TLDR

PCR-based methods for detecting haemosporidian parasites in avian blood have been developed and are considerably more sensitive than traditional microscopy, profoundly influencing avian parasite research. The study aims to introduce a new nested PCR approach that improves detection of these parasites. The new method builds on a previously published PCR protocol by adding a nested amplification step to enhance sensitivity. Compared with existing assays, the new nested PCR detects more Plasmodium spp.

Abstract

Recently, several polymerase chain reaction (PCR)–based methods for detection and genetic identification of haemosporidian parasites in avian blood have been developed. Most of these have considerably higher sensitivity compared with traditional microscope-based examinations of blood smears. These new methods have already had a strong impact on several aspects of research on avian blood parasites. In this study, we present a new nested PCR approach, building on a previously published PCR method, which has significantly improved performance. We compare the new method with some existing assays and show, by sequence-based data, that the higher detection rate is mainly due to superior detection of Plasmodium spp. infections, which often are of low intensity and, therefore, hard to detect with other methods.