Detection of herpes simplex virus in genital specimens by type-specific polymerase chain reaction

Abstract

Viral isolation is the standard method for the detection of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in clinical specimens. This study describes the development of a type-specific polymerase chain reaction (PCR) assay for detection and typing of HSV-1 and HSV-2, and a comparison of its sensitivity with that of isolation in a clinical setting. Specimens from patients presenting with genital ulcers were tested for the presence of HSV by both methods. Oligonucleotide primers were selected to enable type-specific amplification of HSV-1 and HSV-2 DNA. Conditions were optimized to allow detection and typing from a single reaction tube using a multi-primer PCR method. When compared with PCR, the sensitivity of isolation was 67% and the specificity 97%. This protocol allowed rapid, sensitive and accurate detection and typing of HSV with a single PCR assay.

Increasing relative prevalence of HSV-2 infection among men with genital ulcers from a mining community in South Africa

“…Differentiation of HSV-1 and HSV-2 using either PCR or serology based assays have been reported in several studies. [28][29][30] Most recently, real time PCR using the LightCycler (Roche Diagnostics Corp, Indianapolis, IN, USA) has been used to diagnose HSV infections and to type HSV-1 and HSV-2 by melting curve analysis. 31 Our HSV typing system using the Smart Cycler also employed the fluorescence resonance energy transfer (FRET) technology to detect PCR products; in addition, a M-PCR assay using different fluorescent reporter molecules on the TaqMan probes was used to differentiate HSV-1 and HSV-2.…”

Polymerase chain reaction for diagnosis of genital herpes in a genitourinary medicine clinic

“…8 Four previous published studies have compared the performance of PCR with virus culture diagnosis of genital herpes in a routine clinical setting, with consistent findings of superior sensitivity of PCR; compared with PCR, the sensitivity of culture ranged between 67% and 81%. [9][10][11][12] Cullen et al compared PCR and culture, using combined culture and PCR as a "gold standard" in specimens taken from 112 ulcerative genital lesions. 9 PCR had a sensitivity of 93.2% and a specificity of 100%, compared with respective values of 84.1% and 100% for culture.…”

Typing of herpes simplex virus from human periodontium

“…Previous methods of herpes simplex virus culture confirmation were relatively insensitive and sometimes failed to identify HSV despite strong clinical evidence of HSV infection, especially with decreasing viral titers in later disease stages ( 15). Recently, polymerase chain reaction (PCR)‐based identification has emerged as a rapid, specific and sensitive diagnostic test for HSV ( 14, 16). The present study employed a single PCR method for HSV typing.…”