Detection of herpes simplex virus DNA from genital lesions by in situ hybridization

Abstract

Lesion specimens from 118 episodes of recurrent genital herpes were used to compare herpes simplex virus (HSV) isolation with a direct specimen test for in situ DNA hybridization utilizing a biotinylated probe. The frequency of detection of HSV was similar with both tests; HSV was isolated from 81% of vesicular lesions, 76% of pustules, and 67% of ulcers, while HSV DNA was detected in 77, 76, and 55% of lesions in these stages, respectively. Utilizing both methods, HSV was identified in 91, 94, and 79%, respectively. The sensitivity and specificity of the DNA probe in comparison to standard viral isolation in tissue culture were 92 and 63%, respectively. Seven DNA-positive, viral isolation-negative specimens were obtained from patients who had positive culture confirmation at some time subsequent or prior to enrollment, suggesting that these were true positive results. The sensitivity of the DNA probe was dependent on cellular content of the specimen, and 36 (28%) of the 127 submitted specimens had fewer than 20 nonsuperficial cells. The DNA probe was rapid and convenient; its major disadvantage was the lack of type-specific information. The performance of the probe in lower-prevalence populations and in asymptomatic shedding of HSV remains to be evaluated.

A rapid simple in situ hybridization method for herpes simplex virus employing a synthetic biotin‐labeled oligonucleotide probe: A comparison with immunohistochemical methods for HSV detection

“…Viral cultures have been considered the gold standard for -HSV identification; however, this method typically takes several days to a week, although coupling culture with several antigenic or hybridization detection techniques have been evaluated as an alternative approach for rapid diagnosis (4). Nonisotopic in situ hybridization (ISH) techniques performed directly on tissue and cytologic specimens are an attractive possibility as they require as little as 1-2 hr to perform and may be comparable to viral cultures in sensitivity (5).…”

Optimization of in situ hybridization for detection of viral genomes in cultured cells on 96-microwell plates: a cytomegalovirus model

“…The availability of viral DNA probes has modified considerably the diagnosis of viral infections (3,4,11,15,19,29,33). Our intention was to adapt ISH methodology to 96microwell microplates and especially to apply this methodology to the research of infectious CMV isolated by cell culture, i.e., after inoculation of MRC-5 fibroblasts with biological specimens.…”

Detection of herpes simplex virus by the Kodak SureCell Herpes Test

“…Quicker, if somewhat less sensitive, results can be obtained by various nonculture techniques. These include the cytological examination of lesion cells by Wright-Giemsa stain (Tzanck preparation) (3,17), the use of fluorescein-conjugated HSV antibodies in a direct immunofluorescence test (14), immunoperoxidase staining (6,14), direct enzyme immunoassay (EIA) (7,10), and DNA hybridization (4,8).…”

“…That SC-HSV was found to be more sensitive for vesicular lesions and for first-episode genital HSV infections is not surprising. It has been shown that the ability to detect HSV (antigen) varies over the course of the disease, and viral titers have been found to be highest in the vesicular stages of the disease (6,8,11). Also, first episodes of genital herpes often have higher viral titers and a longer duration of virus shedding (1,6).…”