在这项研究中开发了可用于敏感地检测不同亚科的几种疱疹病毒(HV)物种的共有PCR测定法。设计了含有简并碱基的引物,以扩增α-和γ-HVs的DNA聚合酶(DPOL)基因的区域,和β-HVs的糖蛋白B(gB)基因呈单重形式,非嵌套触地PCR格式。单重触地共有PCR(STC-PCR)用于扩增8个人和24只动物HV的DNA。该测定能够检测α-HV的10-5和β-和γ-HV的10-3的最低DNA稀释度。相比之下,α-的最低检出限为10-5、10-3和10-2,beta-,当使用巢式PCR时,分别为γ-HV。这项研究的结果表明,STC-PCR测定可用于分子调查和新型和已知的HV的临床检测。
Consensus PCR assays that can be used to sensitively detect several herpesvirus (HV) species across the different subfamilies were developed in this study. Primers containing degenerate bases were designed to amplify regions of the DNA polymerase (DPOL) gene of alpha- and gamma-HVs, and the glycoprotein B (gB) gene of beta-HVs in a singleplex, non-nested touchdown PCR format. The singleplex touchdown
consensus PCR (STC-PCR) was used to amplify the DNA of eight human and 24 animal HVs. The assay was able to detect the lowest DNA dilution of 10-5 for alpha-HVs and 10-3 for beta- and gamma-HVs. In comparison, lowest detection limits of 10-5, 10-3, and 10-2 were obtained for alpha-, beta-, and gamma-HVs respectively when a nested PCR was used. The findings in this study suggest that the STC-PCR assays can be employed for the molecular surveys and clinical detection of novel and known HVs.