Abstract: A quantitative PCR assay was developed for specific and sensitive detection of mink enteritis virus. The primer pair targeting a 146 bp fragment of the conserved VP2 gene was selected based on alignment of viral sequences available at GenBank. The analytic sensibility of this assay was 34 copies of plasmid DNA,and it had high repeatability with CV <2% ． The assay was evaluated by testing 80 fecal samples collected from suspicious cases. The results showed that 30 samples were found positive for the virus infection,which was higher than that of routine PCR and electron microscopy. Therefore the new assay provides a useful method for the rapid diagnosis of mink enteritis and the prevention and control of the disease.

Key words:  Fluorescence quantitative PCR, Mink enteritis virus, SYBR Green I

摘要： 根据GenBank 登录的水貂肠炎病毒（ＭＥＶ）ＶＰ２ 蛋白基因的保守序列设计了１ 对特异性引物，经ＰＣＲ 扩增出１４６ ｂｐ 的目的片段，并克隆到ｐＭＤ １８ － Ｔ 载体上，提取重组质粒经ＰＣＲ 及测序鉴定正确后，以１０ 倍梯度稀释质粒作为阳性标准品，绘制荧光定量标准曲线，以此建立一种基于ＳＹＢＲ Green 荧光染料的定量ＰＣＲ 特异性检测水貂肠炎病毒的方法。结果表明：该方法对ＭＥＶ 有很好的特异性，检测灵敏度为３４ 拷贝／ μＬ，且重复性试验变异系数均小于２％ 。临床检测中，在８０ 份样品中检测到３０ 份阳性样品，高于普通ＰＣＲ 和电镜观察方法的检出率。该方法的建立实现了对ＭＥＶ 临床早期快速诊断和定量分析，为毛皮动物ＭＥＶ 的防控提供了可靠的检测方法。

关键词: 水貂肠炎病毒, SYBR Green I, 荧光定量PCR