Detection of herpesvirus cervicovaginitis by a sequential papanicolaou‐immunoperoxidase technique

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Abstract
Herpes simplex virus (HSV) infection constitutes an immediate threat to the neonate of pregnant women who deliver vaginally, and thus requires a rapid, specific means of diagnosis that is easily applicable to cervicovaginal smears. We applied the peroxidase-antiperoxidase technique to variously fixed, previously stained Papanicolaou smears from 60 women with HSV and 18 negative controls using an HSV-2 antibody and either diaminobenzidine (DAB) or aminoethylcarbazol (AEC) as the chromogen and Mayer's, Gill's, or Lillie-Mayer's hematoxylin as the counterstain. With Papanicolaou stain alone, there was adequate cytologic demonstration of either single cells in aggregates (11%), syncytial giant cells (40%), or both (49%) that displayed a ground-glass appearance (68%), discrete nuclear inclusions (5%), or both (28%). With the peroxidase-antiperoxidase technique, 57 of the 60 HSV specimens (95%) stained moderately or strongly positive for HSV-2, as did sections of herpetic encephalitis and esophagitis. There was no false-positive staining in any of the 18 control smears revealing koilocytosis, Chlamydia, or nonspecific vaginitis. Positivity of the immunostain was more vivid and evenly dispersed through-out the cytoplasm with AEC than with DAB, but tended to wash away with alcohol-based counterstaining. In contrast, DAB was more stable, but was positive predominantly at the cell periphery and cytoplasmic processes. Lillie-Mayer's stain provided the best counterstaining for the cytologic visualization of virocytes and accompanying inflammatory and epithelial cells, which revealed a minimal degree of atypia. The fixative used had no influence on the frequency or degree of immunopositivity of virocytes, but wet fixation with 95% alcohol or Carbowax led to less background staining than Spray-Cyte.(ABSTRACT TRUNCATED AT 250 WORDS)