Department of Veterinary Pathology, University of Liverpool, Neston, Cheshire, UK.
BMC Vet Res. 2010 Jun 8;6:33. doi: 10.1186/1746-6148-6-33.
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. 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.
This 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.
The 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.
松鼠痘病毒（SQPV）对红松鼠（Sciurus vulgaris）具有高度致病性，是导致英格兰和威尔士大部分地区该物种局部灭绝的一个重要因素，在这些地区，感染是东部灰松鼠（Sciurus carolinensis）种群中的地方病。尽管巢式 PCR 检测法已成功用于研究 SQPV 的流行病学，但样本处理时间长，且该检测法无法定量。
本项目描述了一种针对 SQPV 的实时 PCR 的设计和优化。与巢式 PCR 的比较表明，实时检测法的灵敏度高一个对数级，能够检测到每毫克组织约 144 个基因组拷贝。
实时 PCR 已用于定量患病和看似健康的红松鼠和灰松鼠组织中的病毒基因组载量，表明患病红松鼠组织中的病毒滴度明显高于甚至在患有皮肤损伤的灰松鼠中发现的滴度。
