应用实时荧光定量PCR相对定量法检测毛皮动物细小病毒载量

摘要

参考GenBank中已登录的CPV-SC-JM VP2基因序列设计引物，选用β-actin作为内参基因，利用SYBR Green Ⅰ实时定量PCR相对定量法，对规模化毛皮动物场的12份粪便样品和5份血液样品进行检测。常规PCR方法检出14份，检出率82％，荧光定量PCR相对定量方法检出17份，检出率100％；粪便中病毒含量明显高于血液中病毒含量，最高可达7000倍；不同毛皮动物粪便样品中病毒含量也存在较大差异。%A SYBR Green Ⅰ relative quantification real-time PCR method was used for detection of parvovirus loads in blood and faeces of using primers targeting the nuecleocide sequence logged in the GenBank .The results showed that the real -time PCR was more sensitive than the conventional PCR respectively with 100% and 82% positive detection rates .The levels of virus loads in faeces were significantly higher than that in the blood ,up to 7000-fold ,it also various largely in different faeces samples from diverse fur animals .