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

Department of Animal Ecology, Lund University, Lund, Skåne, Sweden
Journal of Parasitology (Impact Factor: 1.23). 03/2004; 90(1):191-4. DOI: 10.1645/GE-3221RN
Source: PubMed


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.

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Available from: Jonas Waldenström, Jul 04, 2014
    • "Additionally, DNA was extracted from blood samples using a standard ammonium acetate method. The PCR-based method for detection of haemosporidians (Waldenström et al. 2004) was used for confirming the absence or presence of Plasmodium and Haemoproteus infections, without identifying genetic lineages via sequencing of the obtained PCR products. All samples identified by microscopy as negative were confirmed by PCR-based diagnostics . "

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    • "The genera Plasmodium, Haemoproteus and Leucocytozoon represent a group of vector-borne blood parasites causing a malaria-like disease in birds (e.g. Bensch et al. 2000; Waldenstöm et al. 2004; Bensch and A ˚ kesson 2003; Scheuerlein and Ricklefs 2004; Wood et al. 2007; Jenkins and Owens 2011; Swanson et al. 2014; Zhao et al. 2014). One of the features of such infections is that host species vary substantially in parasite prevalence. "
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    • "DNA from the avian blood samples were extracted in the lab using the standard phenol/chloroform/isoamylalcohol method (Sambrook et al. 2002). Diluted genomic DNA (25 ng/μl) was used as a template in a polymerase chain reaction (PCR) assay for detection of the parasites using nested PCR protocols described by Waldenström et al. (2004). The amplification was evaluated by running 2.5 μl of the final PCR on a 2 % agarose gel. "
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