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.
OBJECTIVE
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.
Department of Virology, Faculty of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.
Mohammad Ali
MA
Khodadoost
Farzaneh
F
Sabahi
Mahmoud Jabbarvand
MJ
Behroz
Mohammad Hassan
MH
Roustai
Horieh
H
Saderi
Samad
S
Amini-Bavil-Olyaee
Mohsen Karimi
MK
Arzenani
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D002522:Chlorocebus aethiops; D003588:Cytopathogenic Effect, Viral; D004279:DNA, Viral / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006367:HeLa Cells; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007492:Iran; D016849:Keratitis, Herpetic / Q000821:virology*; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods; D013666:Tears / Q000821:virology; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics*; D014776:Virus Cultivation / Q000379:methods
D000914:Antibodies, Viral; D004279:DNA, Viral; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1