SCIENTIFIC REPORT

Diagnosis of herpes simplex virus-1 keratitis using Giemsa stain, immunofluorescence assay, and polymerase chain reaction assay on corneal scrapings

S Farhatullah¹, S Kaza², S Athmanathan¹, P Garg², S B Reddy¹ and S Sharma¹

¹ Jhaveri Microbiology Center, Professor Brien Holden Eye Research Center, Hyderabad Eye Research Foundation, LV Prasad Eye Institute, LV Prasad Marg, Banjara Hills, Hyderabad - 500 034, India
² Cornea Services, LV Prasad Eye Institute, LV Prasad Marg, Banjara Hills, Hyderabad - 500 034, India

Correspondence to:
Correspondence to:
Savitri Sharma, MD
Jhaveri Microbiology Center, LV Prasad Eye Institute, LV Prasad Marg, Banjara Hills, Hyderabad - 500 034, India; savitri{at}lvpeye.org

Aims: To evaluate three tests used routinely for the diagnosis of herpes simplex virus (HSV) keratitis.

Methods: Corneal scrapings from 28 patients with clinically typical dendritic corneal ulcer suggestive of HSV keratitis, and 30 patients with clinically non-viral corneal ulcers, were tested by (i) Giemsa stain for multinucleated giant cells, (ii) immunofluorescence assay (IFA) for HSV-1 antigen, and (iii) polymerase chain reaction (PCR) for HSV-1 DNA, by investigators masked to clinical diagnosis. The control subjects were also investigated by smears and cultures for bacteria, fungus, and Acanthamoeba.

Results: The specificity and positive predictive values of all three tests for the diagnosis of HSV keratitis were between 95–100%. The sensitivity of IFA and PCR was 78.6% and 81.2%, respectively, and the difference was not significant; however, their sensitivity and negative predictive value were significantly higher than Giemsa stain.

Conclusions: While a combination of IFA and PCR constitute the choice of tests in clinically suspected cases of HSV keratitis, multinucleated giant cells in Giemsa stain can pre-empt testing by IFA and PCR in otherwise atypical cases of HSV keratitis.

Keywords: herpes simplex virus; keratitis; corneal scrapings

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