用于猪巨细胞病毒检测的PCR系统的开发及其DNA聚合酶基因假定部分序列的测定。
Development of a PCR system for porcine cytomegalovirus detection and determination of the putative partial sequence of its DNA polymerase gene.
作者信息
Widen B F, Lowings J P, Belak S, Banks M
机构信息
Virology Department, Central Veterinary Laboratory/Veterinary Laboratories Agency, New Haw, Addlestone, United Kingdom.
出版信息
Epidemiol Infect. 1999 Aug;123(1):177-80. doi: 10.1017/s0950268899002599.
After PCR amplification with conservative cytomegalovirus primers, a 520 nucleotide putative partial sequence of the DNA polymerase gene of porcine cytomegalovirus (PCMV) was determined. Sequence comparison revealed homology to DNA polymerase genes from various beta herpes viruses and a dendrogram was constructed depicting the relationship of PCMV to other members of the Herpesviridae family. The dendrogram indicates that PCMV is indeed a beta herpes virus that is more closely related to human herpes virus types 6 and 7 than to type 5. To address the difficulties encountered during conventional PCMV detection and characterization a set of nested PCR primers were constructed which generated DNA fragments of 415 and 257 bp from the DNA polymerase gene. The nested PCR system proved specific for PCMV and provided a novel means for the detection of this poorly characterized herpes virus in pig populations, vaccines and in organs used in xenotransplantation.
用保守的巨细胞病毒引物进行PCR扩增后，确定了猪巨细胞病毒（PCMV）DNA聚合酶基因一段520个核苷酸的假定部分序列。序列比较显示与各种β疱疹病毒的DNA聚合酶基因具有同源性，并构建了一个树形图来描述PCMV与疱疹病毒科其他成员的关系。该树形图表明，PCMV确实是一种β疱疹病毒，与人类疱疹病毒6型和7型的关系比与5型更为密切。为了解决传统PCMV检测和鉴定过程中遇到的困难，构建了一组巢式PCR引物，这些引物从DNA聚合酶基因产生了415和257 bp的DNA片段。巢式PCR系统被证明对PCMV具有特异性，并为在猪群、疫苗以及异种移植所用器官中检测这种特征不明的疱疹病毒提供了一种新方法。
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