Baron J M, Rübben A, Grussendorf-Conen E I
Department of Dermatology, RWTH-Aachen, Germany.
J Med Virol. 1996 Aug;49(4):279-82. doi: 10.1002/(SICI)1096-9071(199608)49:4<279::AID-JMV4>3.0.CO;2-0.
The polymerase chain reaction (PCR) enables rapid and sensitive detection of VZV and HSV DNA and its efficiency depends mainly on the choice of the primers. Primers should hybridize to conserved DNA sequences within the viral genomes in order to avoid unreliable amplification due to DNA sequence variation between different strains. The aim of the study was to design and to evaluate a general primer pair which permits fast and reliable detection of HSV and VZV. The genes UL 15 of HSV and UL 42 of VZV share the highest degree of homology within the two genomes. We designed a primer pair (GPHV-RU) which hybridizes to these genes. The genetic variability of amplified sequences from clinical specimens was analyzed by restriction enzyme cleavage analysis and by temperature gradient SSCP analysis (TG-SSCP). PCR with GPHV-RU amplified viral sequences from all analyzed specimens (25 x VZV, 10 x HSV-1, 5 x HSV-2) obtained from patients with clinical evidence of HSV or VZV infection. Restriction enzyme cleavage analysis with Hpa II further permitted reliable distinction between VZV, HSV-1, and HSV-2. Analysis of the heterogeneity of the amplified sequences by restriction enzyme cleavage and by TG-SSCP demonstrated no variability between the analyzed clinical specimens of VZ and of HSV-2 and only one differing TG-SSCP-pattern within the HSV-1 isolates. The results suggest that detection of HSV and VZV using the new primer pair GPHV-RU should give reliable results as the amplified sequences show little genetic variability within clinical isolates of HSV-1/2 and VZV.
聚合酶链反应（PCR）能够快速、灵敏地检测水痘带状疱疹病毒（VZV）和单纯疱疹病毒（HSV）的DNA，其效率主要取决于引物的选择。引物应与病毒基因组内的保守DNA序列杂交，以避免因不同毒株之间的DNA序列变异而导致的不可靠扩增。本研究的目的是设计并评估一对通用引物，用于快速、可靠地检测HSV和VZV。HSV的UL 15基因和VZV的UL 42基因在两个基因组中具有最高程度的同源性。我们设计了一对与这些基因杂交的引物（GPHV-RU）。通过限制性内切酶切割分析和温度梯度单链构象多态性分析（TG-SSCP）对临床标本中扩增序列的遗传变异性进行了分析。使用GPHV-RU进行的PCR扩增了来自所有分析标本（25份VZV、10份HSV-1、5份HSV-2）的病毒序列这些标本取自具有HSV或VZV感染临床证据的患者。用Hpa II进行的限制性内切酶切割分析进一步能够可靠地区分VZV、HSV-1和HSV-2。通过限制性内切酶切割和TG-SSCP对扩增序列的异质性分析表明，在分析的VZ和HSV-2临床标本之间没有变异性，在HSV-1分离株中只有一种不同的TG-SSCP模式。结果表明，使用新引物对GPHV-RU检测HSV和VZV应该能得到可靠的结果，因为扩增序列在HSV-1/2和VZV的临床分离株中显示出很小的遗传变异性。
