Abstract
BackgroundSquirrel poxvirus (SQPV) is highly pathogenic to red squirrels (Sciurus vulgaris), and is a significant contributing factor to the local extinction of the species in most parts of England and Wales, where infection is endemic in Eastern grey squirrel (Sciurus carolinensis) populations. Although a nested PCR assay has been used successfully to study the epidemiology of SQPV, samples have a long processing time and the assay is not quantifiable.ResultsThis project describes the design and optimization of a real-time PCR for SQPV. Comparison with the nested PCR showed the real-time assay to be more sensitive by one log and able to detect approximately 144 genome copies per mg of tissue.ConclusionsThe real-time PCR has been used to quantify viral genome load in tissues from diseased and apparently healthy red and grey squirrels, and suggests that the titre of virus in tissues from diseased red squirrels is considerably higher than that found even in a grey squirrel with cutaneous lesions.
Highlights
Squirrel poxvirus (SQPV) is highly pathogenic to red squirrels (Sciurus vulgaris), and is a significant contributing factor to the local extinction of the species in most parts of England and Wales, where infection is endemic in Eastern grey squirrel (Sciurus carolinensis) populations
We describe a nested and real-time polymerase chain reaction (PCR) assay, both of which were able to detect poxviral DNA from tissues from red and grey squirrels naturally infected with squirrel pox virus
Realtime PCR quantification suggests detection is sensitive to approximately 144 genome equivalents per mg tissue which is comparable with other real-time poxviral assays [15]
Summary
Squirrel poxvirus (SQPV) is highly pathogenic to red squirrels (Sciurus vulgaris), and is a significant contributing factor to the local extinction of the species in most parts of England and Wales, where infection is endemic in Eastern grey squirrel (Sciurus carolinensis) populations. In the UK, squirrelpox virus (SQPV) is known to infect two species of squirrel: the native red squirrel (Sciurus vulgaris) and the introduced Eastern grey squirrel (Sciurus carolinensis) [3]. The disease is characterized by a severe multifocal ulcerative and exudative dermatitis with hemorrhagic scabs which is predominantly fatal if untreated. Such lesions appear primarily on the eyelids, nose and lips, with later spread to the digits, inguinal area and ventral skin of the body [4,5,6,7]. In contrast to red squirrels, grey squirrels generally do not show
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