Donofrio G, Cavirani S, Flammini C F, Scatozza F
Istituto di Malattie Infettive Profilassi e Polizia Veterinaria, Facoltà di Medicina Veterinaria, Università di Parma, Italy.
Vet Res Commun. 2000 Sep;24(6):411-22. doi: 10.1023/a:1006478317972.
A new BHV-4 (bovine herpesvirus 4) isolated from a case of bovine interdigital dermatitis was characterized by PCR and restriction enzyme analysis. To determine whether the new isolate (PR/1) belonged to BHV-4, DNA from infected cells was specifically amplified by PCR. We used a set of primers spanning a large 2.571 kb conserved region of the BHV-4 genome, including the 3' end of ORF1 (homologous to the EBV BVRF1 gene), ORF2 (homologous to the EBV BXRF1 gene), ORF3 (TK gene) and ORF4 (gH gene) 5' end, respectively. The identity of the amplified product was confirmed by HindIII restriction enzyme digestion and Southern hybridization. No product was observed from the DNA of other bovine herpesviruses tested. The restriction patterns of the PR/ 1 genome compared to DN 599, MOVAR 33/63 and LVR BHV-4 reference strains showed two kinds of differences, either related or not related to the prDNA (polyrepetitive DNA). Taken together, these data show that PR/ 1 is a new BHV-4. We would consider that the present report provides a scheme of work for diagnosis and typing of BHV-4 isolates.
从一例牛趾间皮炎病例中分离出的一株新的牛疱疹病毒4型（BHV - 4），通过聚合酶链反应（PCR）和限制性内切酶分析进行了鉴定。为确定新分离株（PR/1）是否属于BHV - 4，采用PCR对感染细胞的DNA进行特异性扩增。我们使用了一组引物，该引物跨越BHV - 4基因组一个2.571 kb的大保守区域，分别包括ORF1（与EBV BVRF1基因同源）的3'端、ORF2（与EBV BXRF1基因同源）、ORF3（胸苷激酶基因，TK基因）和ORF4（糖蛋白H基因，gH基因）的5'端。通过HindIII限制性内切酶消化和Southern杂交确认扩增产物的一致性。在所检测的其他牛疱疹病毒的DNA中未观察到产物。与DN 599、MOVAR 33/63和LVR BHV - 4参考毒株相比，PR/1基因组的限制性图谱显示出两种差异，这两种差异与前病毒DNA（多重复DNA）相关或不相关。综上所述，这些数据表明PR/1是一株新的BHV - 4。我们认为本报告为BHV - 4分离株的诊断和分型提供了一个工作方案。
