摘要
根据GenBank中Ⅰ型鸭病毒性肝炎病毒(DHV-Ⅰ)的基因序列设计、合成引物,以山东、江苏、广东等地分离株的RNA为模板进行RT-PCR扩增,得到预期大小的目的片段。将山东分离株扩增的目的片段克隆到pGM-T载体,酶切鉴定得到阳性重组质粒。对重组质粒进行序列测定,与参考毒株R85952的序列比较发现,山东分离株与参考序列的同源性为97.4%。试验结果表明,所建立的RT-PCR方法最低模板检出量为100 pg,且该方法只能特异地检测出DHV,不能检出鸭瘟病毒、小鹅瘟病毒、鹅副粘病毒和番鸭细小病毒等。自然感染病料的检测分离结果表明,该方法既可用于DHV-Ⅰ分离株的快速鉴定,也可用于临床感染病例的检测与诊断。
A RT-PCR method for detecting the duck hepatitis virusⅠ type was established, A specific fragment was amplified with a pair of primers designed and synthesized based on the gene sequence in GenBank. The fragment was cloned into pGM-T vector, and the positive recombine plasmid was selected and characterized by NotⅠ enzyme digestion.The recombine plasmids were sequenced and compared with pulished sequence, and the homology of Shandong isolated stain was 97.4%. The sensitivity and specificity of the RT-PCR method were very high, which could detect 100 pg RNA of the duck hepatitis virus, but couldn＇t detect duck plague virus, goose parvovirus, goose paramyxoviridae and muscovy duck parvovirus. So the method can be applied for the fastly diagnosis of duck hepatitis virus Ⅰ type and clinical duck hepatitis virus infectious samples.
出处
《广东农业科学》
CAS
CSCD
2007年第7期81-84,共4页
Guangdong Agricultural Sciences
基金
山东省自然基金项目(Z2006D06)
关键词
鸭病毒性肝炎病毒
Ⅰ型
RT-PCR
检测
duck hepatitis virus
Ⅰ type
reverse transcriptional polymerase chain reaction
detection
