Jm. Baron et al., EVALUATION OF A NEW GENERAL PRIMER PAIR FOR RAPID DETECTION AND DIFFERENTIATION OF HSV-1, HSV-2, AND VZV BY POLYMERASE CHAIN-REACTION, Journal of medical virology, 49(4), 1996, pp. 279-282
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.