Detection and analysis of diverse herpes-viral species by consensus primer PCR
Detection and analysis of diverse herpes-viral species by consensus primer PCR
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DOI：
10.1128/jcm.34.7.1666-1671.1996
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发表时间：
1996-07-01
影响因子：
9.4
通讯作者：
Rose, TM
中科院分区：
文献类型：
--
作者：
VanDevanter, DR;Warrener, P;Rose, TM
开发了扩增疱疹病毒DNA指导的DNA聚合酶（EC 2.7.7.7）的一个区域并以嵌套形式使用简并引物的共有引物PCR方法。引物被设计成靶向编码覆盖约800 bp区域的高度保守氨基酸基序的序列。该试验应用于22种疱疹病毒（8人和14动物病毒），获得21的22种病毒的PCR产物。在这个过程中，获得了14个以前未报道的疱疹病毒DNA聚合酶的氨基酸编码序列，包括人类疱疹病毒7和8的区域。在本研究中回收的50至60个氨基酸编码序列被确定为对于所研究的每个病毒物种是独特的，当研究时，单个物种的菌株之间的序列变异非常小。（疱疹病毒使用1、2、3、4、5和6 B）可以在等于或低于100基因组当量/100 ng载体DNA的水平下检测到，这些数据表明，针对疱疹病毒DNA聚合酶的共有引物PCR可能被证明在检测和鉴定已知的疱疹病毒中是有用的。临床样本和新疱疹病毒基因组的初步表征。
A consensus primer PCR method which amplifies a region of herpesviral DNA-directed DNA polymerase (EC 2.7.7.7) and which uses degenerate primers in a nested format was developed, Primers were designed to target sequences coding for highly conserved amino acid motifs covering a region of approximately 800 bp. The assay was applied to 22 species of herpesviruses (8 human and 14 animal viruses), with PCR products obtained for 21 of 22 viruses. In the process, 14 previously unreported amino acid-coding sequences from herpesviral DNA polymerases were obtained, including regions of human herpesviruses 7 and 8. The 50 to 60 amino acid-coding sequences recovered in the present study were determined to be unique to each viral species studied, with very little sequence variation between strains of a single species when studied, Template dilution studies in the presence of human carrier DNA demonstrated that six human herpesviruses (herpesviruses uses 1, 2, 3, 4, 5, and 6B) could be detected at levels at or below 100 genome equivalents per 100 ng of carrier DNA, These data suggest that consensus primer PCR targeted to herpesviral DNA polymerase may prove to be useful in the detection and identification of known herpesviruses in clinical samples and the initial characterization of new herpesviral genomes.