EVALUATION OF A NEW GENERAL PRIMER PAIR FOR RAPID DETECTION AND DIFFERENTIATION OF HSV-1, HSV-2, AND VZV BY POLYMERASE CHAIN-REACTION

The polymerase chain reaction (PCR) enables rapid and sensitive detect ion of VZV and HSV DNA and its efficiency depends mainly on the choice of the primers. Primers should hybridize to conserved DNA sequences w ithin the viral genomes in order to avoid unreliable amplification due to DNA sequence variation between different strains. The aim of the s tudy 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 an d UL 42 of VZV share the highest degree of homology within the two gen omes. We designed a primer pair (GPHV-RU) which hybridizes to these ge nes. The genetic variability of amplified sequences from clinical spec imens was analyzed by restriction enzyme cleavage analysis and by temp erature gradient SSCP analysis (TG-SSCP). PCR with GPHV-RU amplified v iral sequences from all analyzed specimens (25xVZV, 10xHSV-1, 5xHSV-2) obtained from patients with clinical evidence of HSV or VZV infection . Restriction enzyme cleavage analysis with Hpa II further permitted r eliable distinction between VZV, HSV-1, and HSV-2. Analysis of the het erogeneity of the amplified sequences by restriction enzyme cleavage a nd by TG-SSCP demonstrated no variability between the analyzed clinica l specimens of VZV 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 cli nical isolates of HSV-1/2 and VZV. (C) 1996 Wiley-Liss, Inc.