Original article
Detection and genotyping of herpes simplex virus types 1 and 2 by polymerase chain reaction

https://doi.org/10.1016/S0890-8508(95)91508-7Get rights and content

A simple and rapid polymerase chain reaction (PCR) procedure was developed for simultaneous detection and typing of herpes simplex virus (HSV) types 1 and 2. It was possible to detect and type HSV using two primers pairs in a simultaneous double PCR reaction, where the type of HSV present was determined on the basis of an ethidium-bromide-stained band after agarose gel electrophoresis. This PCR assay was tested on about 500 clinical specimens.

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    The conditions for both reactions were as follows: incubation at 50 °C for 2 min, denaturation of DNA at 95 °C for 5 min, followed by 40 cycles of 94 °C for 1 min, 45 s at 58 °C for annealing, an extension step at 72 °C for 30 s, and a final extension step at 72 °C for 10 min. The primers HSV-1a (5′-CCCTGTCTCGCGCGAGCCAC-3′) and HSV-1b (5′-TCAGCCACCCATACGCGTAA-3′), which amplify a fragment of 142 bp [19], were used to detect DNA from HSV-1, while the primers HSV-2 (A) (5′-GGACGAGGCGCCAAAGCACACG-3′) and HSV-2 (B) (5′-TCCGTCCAGTCGTTTATCTTCAC-3′), which generate a product of 270 bp [20], were used for HSV-2. The products of PCRs underwent vertical electrophoresis on polyacrylamide gel at 8% [16], with subsequent revelation by silver [18].

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