Rapid detection of herpes simplex virus (HSV) antigen in human ocular infections.

The new HERPCHEK (Dupont, No. Billerica, MA) enzyme immunosorbent assay (EIA) was used in a double-blind clinical study for rapid and specific detection of ocular herpes simplex virus (HSV) infection. This 4-hour assay can be used to demonstrate conclusively the presence of HSV antigen without culture and thereby rapidly differentiate between HSV and other clinically similar ocular infectious diseases. Ocular samples were collected from 180 individuals including 30 patients with acute HSV, 90 with latent HSV (ie, currently asymptomatic but with a positive history), 11 with acute or latent varicella zoster virus, 30 with nonherpetic infections (due to adenovirus, Acanthamoeba or bacteria), and 19 normal controls. A clinical diagnosis was made by one of us (DPL) and duplicate tear-film samples obtained by swabbing the conjunctival cul-de-sac and cornea. Coded samples were tested by routine viral culture on Vero cell monolayers and also were run independently in the HERPCHEK test. During active HSV infection, the HERPCHEK correlated 100% with clinical diagnosis, and virus culture correlated 90% with clinical diagnosis. In all latent HSV ocular infections, other nonherpetic ocular infections and normal samples, both the HERPCHEK and culture assays were negative.

Topical human fibroblast interferon for acute adenoviral conjunctivitis.

We conducted a prospective, randomized, double-blind, placebo-controlled clinical trial investigating the use of topical human fibroblast interferon (HuIFN-beta) 7.5 x 10(5) IU/ml, one drop 5 times daily, in the treatment of acute epidemic conjunctivitis. Of 50 patients who were initially enrolled, tear-film cultures for adenovirus were positive from 26% for type 8, 12, or 19. Based upon a quantifiable conjunctivitis severity score of 37 patients evaluated after approximately 1 week of therapy, analysis of covariance showed a statistically significant greater improvement with topical HuIFN-beta compared with placebo for affected left eyes (P = 0.02) but not for right eyes (P = 0.5), an effect that could not be adequately explained. We suggest that a higher dosage of topical HuIFN-beta may prove useful in the control of more severely affected cases. This trial provides guidelines for future investigations of interferon in the treatment and prophylaxis of adenoviral keratoconjunctivitis.

Detection of herpes simplex virus induced polypeptides in rabbit trigeminal ganglia.

The objective of this study was to follow herpes simplex virus type 1 (HSV-1) genome expression in rabbit trigeminal ganglia during primary, latent, and artificially reactivated infections using monospecific antiserum to representatives of the sequentially produced alpha, beta, and gamma group polypeptides. Rabbits with or without electrode implants were inoculated with 10(5) plaque forming units of McKrae strain HSV-1 following scarification and monitored by daily preocular tear film culture. Animals were killed during primary, latent, and artificially reactivated infections. Representative trigeminal ganglion sections from each animal were reacted with either preimmune rabbit serum, anti-HSV-1, anti-ICP-4, anti-ICP-8, or anti-ICP-5 sera. During primary infection, staining was evident in 30-40% of the ganglion cell nuclei with all antisera. During latency, staining with anti-ICP-4 was detected in 14 out of 18 ganglia tested; 10-15% of the ganglion cell nuclei per section exhibited positive staining. A +/- staining pattern with anti-ICP-8 was obtained in 5 out of 14 of the ganglia tested; 1-3% of the nuclei per section exhibited this staining pattern. Staining with anti-HSV-1 and anti-ICP-5 was not detected. During induced reactivation, staining of ganglion cell nuclei with all antisera was observed, but the number of cells staining per section was decreased compared to that observed during primary disease.

Detection of an immediate early herpes simplex virus type 1 polypeptide in trigeminal ganglia from latently infected animals.

In this study, trigeminal sensory ganglia from animals with acute herpes simplex virus, type 1 (HSV-1) infection were compared to those with a latent infection for the expression of HSV-specific antigens. By the indirect immunofluorescence assay, antisera to an immediate early polypeptide of molecular weight 175,000, designated VP175 or ICP4, and a hyperimmune antiserum to HSV-1 were used to determine whether early viral polypeptides were being expressed in neurons during the latent stage of infection. All 17 ganglia from animals with acute infection (sacrificed 3 to 12 days postinfection) exhibited positive staining when treated either with anti-HSV-1 or with anti-VP175. Forty of 42 ganglia from animals sacrificed during the latent stage of infection (22 to 200 days postinfection) exhibited immunofluorescent staining when treated with anti-VP175. The staining appeared to be similar to that observed in ganglia from acutely infected animals stained with anti-VP175, except that the number and distribution of stained cells were markedly reduced. No immunofluorescence was observed in ganglia from noninfected control animals when stained with anti-VP175 or anti-HSV-1, or when ganglia from latently infected animals were stained with anti-HSV-1 or preimmune serum.