PMC31434_01
Clinical and virological investigations: A. Slit lamp biomicroscopy under optical section of the right eye showing a solitary paracentral, well circumscribed, large, full thickness stromal infiltrate, collapsed anterior chamber and a central area of perforation. X 16. B. Giemsa stained corneal scraping showing a multinucleated giant cell with characteristic molding of the nuclei. X 1250. C. Conjunctival scraping from lower palpebral conjunctiva showing epithelial cells positive for HSV-1 antigen (Indirect immunoperoxidase, X 500). D. Corneal scraping showing many cells positive for HSV-1 antigen (Indirect immunoperoxidase, X 500).
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Anterior chamber was shallow (Fig. 1A). However, Giemsa stained smear showed the presence of multinucleated giant cells, with characteristic molding of nuclei (Fig. 1B), suggesting infectious keratitis, probably of a viral etiology. Scrapings obtained from the lower palpebral conjunctiva, on the following day (corneal scrapings could not be collected due to the application of tissue adhesive and a bandage contact lens), was positive for HSV-1 antigen (Fig. 1C) by an immunoperoxidase assay. HSV-1 antigen was detected by an immunoperoxidase assay (Fig. 1D) in the repeat corneal scrapings while bacterial and viral cultures (shell vial and conventional tube cultures for HSV-1) were negative.
A. Corneal tissue section showing absence of HSV-1 antigen in the epithelium (Negative control, Immunohistochemistry, X 500). B. Corneal tissue section showing presence of HSV-1 antigen in the epithelium (Immunohistochemistry, X 500). C. Corneal tissue section showing a multinucleated giant cell (arrow) (PAS, X 500). D. Agarose gel electrophoresis stained with ethidium bromide showing PCR amplified products using primers for a 179 bp sequence of the DNA polymerase gene specific for HSV-1/2. Lane 1: Negative control (NC); Lane 2: Patient's corneal tissue (VI 65/99); Lane 3: Positive control (PC); Lane 4: Molecular weight markers (MW).
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HSV-1 antigen by immunohistochemistry (Fig. 2A, B) and for HSV DNA by PCR (Fig. 2D). Histopathological examination of the formalin fixed corneal tissue revealed polymorpho nuclear neutrophils admixed with mononuclear cells, multinucleated giant cells in H & E and PAS (Fig. 2C) stained sections.