Abstract
Background Avipoxvirus sp. is a significant threat to endemic bird populations on several groups of islands worldwide, including Hawaìi, the Galapagos Islands, and the Canary Islands. Accurate identification and genotyping of Avipoxvirus is critical to the study of this disease and how it interacts with other pathogens, but currently available methods rely on invasive sampling of pox-like lesions and may be especially harmful in smaller birds.Methodology/Principal FindingsHere, we present a nested TaqMan Real-Time PCR for the detection of the Avipoxvirus 4b core protein gene in archived blood samples from Hawaiian birds. The method was successful in amplifying Avipoxvirus DNA from packed blood cells of one of seven Hawaiian honeycreepers with confirmed Avipoxvirus infections and 13 of 28 Hawaìi `amakihi (Hemignathus virens) with suspected Avipoxvirus infections based on the presence of pox-like lesions. Mixed genotype infections have not previously been documented in Hawaìi but were observed in two individuals in this study.Conclusions/SignificanceWe anticipate that this method will be applicable to other closely related strains of Avipoxvirus and will become an important and useful tool in global studies of the epidemiology of Avipoxvirus.
Highlights
Avian pox virus (Avipoxvirus sp.) has caused extensive morbidity and mortality in the native Hawaiian avifauna [1,2] and currently threatens endemic birds in the Galapagos and Canary Islands [3,4,5,6]
The results from a Taqman Real-Time PCR completed on a serial dilution of first reaction products from a known positive sample are shown in Figure 1 and Table 1
In real-time diagnostic assays, a positive or negative result is often determined by the cycle number at which signal from a sample crosses a baseline threshold (Ct)
Summary
Avian pox virus (Avipoxvirus sp.) has caused extensive morbidity and mortality in the native Hawaiian avifauna [1,2] and currently threatens endemic birds in the Galapagos and Canary Islands [3,4,5,6]. They differ in virulence, and have been shown to cause mortality among naıve Hawaiiamakihi (Hemignathus virens), hereafteramakihi, under experimental conditions [8]. One of these variants has been shown to have a very close phylogenetic relationship with canarypox, as have Avipoxvirus variants present in the Galapagos Islands [4]. Accurate identification and genotyping of Avipoxvirus is critical to the study of this disease and how it interacts with other pathogens, but currently available methods rely on invasive sampling of pox-like lesions and may be especially harmful in smaller birds
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