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1.  Collection of corneal impression cytology directly on a sterile glass slide for the detection of viral antigen: An inexpensive and simple technique for the diagnosis of HSV epithelial keratitis – A pilot study 
BMC Ophthalmology  2001;1:3.
Background
Herpes simplex keratitis (HSK) is a sight threatening ocular infection and occurs worldwide. A prompt laboratory diagnosis is often very useful. Conventional virology techniques are often expensive and time consuming. We describe here a highly economical, simple, rapid and sensitive technique for the collection of impression cytology, for the laboratory diagnosis of HSK.
Methods
Fifteen patients with a clinical diagnosis of HSK (either dendritic or geographic ulcers) and five patients with other corneal infections (Mycotic keratitis, n = 3, Bacterial keratitis, n = 2) were included in the study. Corneal impression cytology specimens were collected using a sterile glass slide with polished edges instead of a membrane, by pressing the surface of one end of the slide firmly, but gently on the corneal lesion. Additionally, corneal scrapings were collected following the impression cytology procedure. Impression cytology and corneal scrapings were stained by an immunoperoxidase or immunofluorescence assay for the detection of HSV-1 antigen using a polyclonal antibody to HSV-1. Corneal scrapings were processed for viral cultures by employing a shell vial assay.
Results
This simple technique allowed the collection of adequate corneal epithelial cells for the detection of HSV-1 antigen in a majority of the patients. HSV-1 antigen was detected in 12/15 (80%) cases while virus was isolated from 5/15 (33.3%) patients with HSK. All the patients with a clinical diagnosis of HSK (n = 15) were confirmed by virological investigations (viral antigen detection and/or viral cultures). HSV-1 antigen was detected in the impression cytology smears and corneal scrapings in 11/15 (73.3%) and 12/15 (80%) of the patients, respectively (P = 1.00). None of the patients in the control group were positive for viral antigen or virus isolation. Minimal background staining was seen in impression cytology smears, while there was some background staining in corneal scrapings stained by the immunoassays.
Conclusions
Collection of impression cytology on a sterile glass slide is a simple, rapid and inexpensive technique for the diagnosis of HSK. Immunological techniques applied on such smears provide virological results within 2-5 hours. This technique could be modified for use in the diagnosis of other external eye diseases, which needs further evaluation.
doi:10.1186/1471-2415-1-3
PMCID: PMC57753  PMID: 11592921
2.  Herpes simplex virus bullous keratitis misdiagnosed as a case of pseudophakic bullous keratopathy with secondary glaucoma: an unusual presentation 
BMC Ophthalmology  2001;1:2.
Purpose
To report an unusual case of herpetic bullous keratitis misdiagnosed as a case of pseudophakic bullous keratopathy with secondary glaucoma.
Results
A retrospective analysis of the case record of a 60-year-old man who had earlier undergone bilateral cataract surgery, was done. He presented with a complaint of decrease in vision in the right eye of 20 days duration. On examination, cornea showed epithelial bullae all over the surface with stromal and epithelial edema. Intraocular pressure was 30 mm of Hg in RE. He was treated with anti-glaucoma medications. Two dendritic lesions were seen in the cornea during a subsequent visit four days later. Virological investigations confirmed a diagnosis of Herpes simplex keratitis. He was treated with topical acyclovir.
Conclusions
This case highlights the fact that herpes simplex keratitis can present initially as a more diffuse corneal stromal and epithelial edema with epithelial bullae mimicking bullous keratopathy. Herpetic bullous keratitis, although unusual, should be considered in the differential diagnosis under such circumstances.
doi:10.1186/1471-2415-1-2
PMCID: PMC35284  PMID: 11472638
3.  Atypical Herpes simplex keratitis (HSK) presenting as a perforated corneal ulcer with a large infiltrate in a contact lens wearer: multinucleated giant cells in the Giemsa smear offered a clue to the diagnosis 
BMC Ophthalmology  2001;1:1.
Purpose
To report a case of atypical herpes simplex keratitis initially diagnosed as bacterial keratitis, in a contact lens wearer.
Results
Case report of an 18-year-old woman using contact lenses who presented with pain, redness and gradual decrease in vision in the right eye. Examination revealed a paracentral large stromal infiltrate with a central 2-mm perforation. Corneal and conjunctival scrapings were collected for microbiological investigations. Corneal tissue was obtained following penetrating keratoplasty. Corneal scraping revealed no microorganisms. Giemsa stained smear showed multinucleated giant cells. Conjunctival, corneal scrapings and tissue were positive for herpes simplex virus - 1 (HSV) antigen. Corneal tissue was positive for HSV DNA by PCR.
Conclusions
Atypical HSV keratitis can occur in contact lens wearers. A simple investigation like Giemsa stain may offer a clue to the diagnosis.
doi:10.1186/1471-2415-1-1
PMCID: PMC31434  PMID: 11325340

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