Volume 48, Number 4, 328-332, DOI: 10.1007/s10384-004-0081-z

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Japanese Ophthalmological Society

Study of a Polymerase Chain Reaction-based Method for Detection of Herpes Simplex Virus Type 1 DNA among Iranian Patients with Ocular Herpetic Keratitis Infection

Mohammad Ali Khodadoost, Farzaneh Sabahi, Mahmoud Jabbarvand Behroz, Mohammad Hassan Roustai, Horieh Saderi, Samad Amini-Bavil-Olyaee and Mohsen Karimi Arzenani

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Abstract

Purpose  

To study the presence of the herpes simplex virus type 1 (HSV-1) glycoprotein D gene in tear films of Iranian patients with herpetic keratitis.

Methods  

Twenty-five tear film and eye swab specimens from 25 herpetic keratitis patients and 10 specimens from 10 healthy volunteers were collected in the Farabi Eye Hospital, Tehran, Iran. HSV-1 DNA was detected by using the nested polymerase chain reaction (nPCR) method. Viral isolation was done using conventional viral techniques. A monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) was used for confirmation of positive cytopathic effect cell culture. The results of a diagnosis by an ophthalmologist team were compared with those of nPCR.

Results  

HSV-1 DNA was identified in tear films of 88% (23/25) of suspected herpetic keratitis patients. All healthy controls (100%) had negative PCR results. HSV-1 was isolated in cell culture and confirmed by ELISA in 12% (3/25) of herpetic keratitis patients who had epithelial keratitis. The kappa value showed a high level of agreement between ophthalmologist team diagnosis and the PCR results (kappa = 0.86, P < 0.0001).

Conclusions  

nPCR is a sensitive, rapid, and powerful tool for detection of HSV-1 DNA in tear films of ocular herpetic keratitis patients and can serve as a supplemental method for diagnosis of herpetic keratitis infection. Jpn J Ophthalmol 2004;48:328–332 © Japanese Ophthalmological Society 2004

Key words  herpes simplex virus type 1 - herpetic keratitis infection - human tear films - nested polymerase chain reaction - viral culture

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