羊传染性脓疱诊断方法的建立

利用MDBK细胞从青海某养殖户发病羊唇部痂皮中分离获得一株病毒,通过临床症状、PCR检测以及组织病理学观察等方法对该株病毒进行了鉴定,证实分离株为羊传染性脓疱病毒(Orf Virus,ORFV),命名为ORFV-QHMH01。参照GenBank中ORFV F1L基因的核苷酸序列,设计合成一对特异性引物,对ORFV-QHMH01株F1L基因进行了克隆、测序,并推导出其氨基酸序列。序列分析表明,分离株F1L基因片段长987bp,编码328个氨基酸,与AF097215、AY040081、AY040082、AY040083、AY040084、AY040085、FJ808075、HM133903、HQ221964、JQ271535株的核苷酸序列同源性分别为:96.7%,95.6%,96.1%,96.2%,97.5%,97.2%,96.5%,48.6%,95.3%,98.3%。系统发育进化树结果表明,ORFV-QHMH01分离株与参考毒株JQ271535和AY040084的亲缘关系较近,这表明不同地区分离株的F1L基因具有较好的保守性。

Establishing on Diagnosis Method of Orf Virus

An Orf virus strain(ORFV-QHMH01) was isolated from scab materials of lambs with clinical score mouth symptom by passaging in MDBK cells.The virus was identified as Orf virus and named ORFV-QHMH01 based on clinical symptoms,PCR and pathohistology observation.A pair of primers derived from the F1L gene of Orf virus(ORFV) was designed according to the published sequence in GenBank.The F1L gene was cloned and sequenced,then the amino acids sequence of F1L protein was deduced.The F1L gene contained 987bp,which encoded a protein of 328 amino acids.The F1L gene of ORFV-QHMH01 strain shared a nucleotide identity of : 96.7%,95.6%,96.1%,96.2%,97.5%,97.2%,96.5%,48.6%,95.3%,98.3% with those of strain AF097215,AY040081,AY040082,AY040083,AY040084,AY040085,FJ808075,HM133903,HQ221964,JQ271535 respectively.The isolated ORFV-QHMH01 has a closer relationship with virus J Q 271535 and AY040084.The results of the phylogenetic tree analysis indicated the F1L gene of different strains has a good conservative.