Deregt D, Jordan L T, van Drunen Littel-van den Hurk S, Masri S A, Tessaro S V, Gilbert S A
Animal Diseases Research Institute, Canadian Food Inspection Agency, Lethbridge, AB, Canada.
Am J Vet Res. 2000 Dec;61(12):1614-8. doi: 10.2460/ajvr.2000.61.1614.
To determine whether a herpesvirus isolated from the semen of a North American elk was related to bovine herpesvirus 1 (BHV-1).
Semen from 1 healthy bull elk and 2 subtypes of BHV-1 (BHV-1.1 and BHV-1.2).
A virus with cytopathic and electron microscopic characteristics consistent with an alpha-herpesvirus was isolated from elk semen, using fetal bovine kidney cells. Cross-neutralization assays were performed with antisera against BHV-1 and the elk herpesvirus (EIkHV). Restriction endonuclease digests of EIkHV DNA were compared with digests of BHV-1.1 and BHV-1.2 DNA. A portion of the ElkHV DNA polymerase gene was amplified with consensus primers by use of the polymerase chain reaction and sequenced. Sequence was compared with known sequences of other herpesviruses. An immunoperoxidase monolayer assay was used to determine reactivities of 22 BHV-1-specific monoclonal antibodies (mAb) against ElkHV. In vitro neutralizing activities of the reactive mAb were determined by use of a microneutralization assay.
Results of cross-neutralization assays indicated that ElkHV was serologically related to BHV-1. Endonuclease digestion of ElkHV DNA generated fragments that were distinct from those of BHV-1. Nucleotide sequencing confirmed that ElkHV is an alphaherpesvirus closely related to but distinct from BHV-1. Six of 22 BHV-1-specific mAb reacted against ElkHV; 2 of these 6 also neutralized in vitro infectivity of ElkHV.
ElkHV is antigenically and genetically distinguishable from BHV-1. However, the viruses are serologically related and share at least 6 antigenic determinants, one of which is a major neutralizing determinant.
确定从一头北美麋鹿精液中分离出的一种疱疹病毒是否与牛疱疹病毒1型(BHV - 1)相关。
来自1头健康雄性麋鹿的精液以及BHV - 1的2个亚型(BHV - 1.1和BHV - 1.2)。
使用胎牛肾细胞从麋鹿精液中分离出一种具有与α疱疹病毒一致的细胞病变和电子显微镜特征的病毒。用抗BHV - 1和麋鹿疱疹病毒(EIkHV)的抗血清进行交叉中和试验。将EIkHV DNA的限制性内切酶消化产物与BHV - 1.1和BHV - 1.2 DNA的消化产物进行比较。使用聚合酶链反应(PCR),用共有引物扩增EIkHV DNA聚合酶基因的一部分并进行测序。将序列与其他疱疹病毒的已知序列进行比较。使用免疫过氧化物酶单层试验来确定22种BHV - 1特异性单克隆抗体(mAb)对EIkHV的反应性。通过微量中和试验确定反应性mAb的体外中和活性。
交叉中和试验结果表明EIkHV与BHV - 1存在血清学关系。EIkHV DNA的内切酶消化产生的片段与BHV - 1的片段不同。核苷酸测序证实EIkHV是一种与BHV - 1密切相关但又不同的α疱疹病毒。22种BHV - 1特异性mAb中有6种对EIkHV有反应;这6种中的2种也能在体外中和EIkHV的感染性。
EIkHV在抗原性和基因上与BHV - 1可区分。然而,这两种病毒存在血清学关系且至少共享6个抗原决定簇,其中一个是主要中和决定簇。