Clinical application of real-time polymerase chain reaction for diagnosis of herpetic diseases of the anterior segment of the eye

Jpn J Ophthalmol. 2008 Jan-Feb;52(1):24-31. doi: 10.1007/s10384-007-0485-7. Epub 2008 Mar 28.


Purpose: To assess the value of quantification of herpes simplex virus (HSV) DNA for the differential diagnosis of herpetic diseases of the anterior segment of the eye.

Methods: One hundred forty-four samples from 90 patients with ocular inflammatory diseases were examined for HSV DNA by real-time polymerase chain reaction (PCR) with primers set for the consensus sequence of HSV-1/2 DNA polymerase. The samples included corneal epithelial scrapings, tear fluid (200microl of eye wash), and aqueous humor.

Results: In cases of typical herpetic epithelial keratitis, a large number of copies of HSV DNA were detected (mean, 1.0 x 10(7) copies in epithelial scrapings and 3.5 x 10(5) copies in tear fluid). In atypical epithelial keratitis cases, a smaller number of HSV DNA copies were detected. In stromal keratitis cases, the number of copies of HSV DNA in the tear fluid (mean: 4.7 x 10(2) copies) was significantly smaller than in cases of epithelial keratitis. In the aqueous humor, the number of copies was small in endotheliitis cases (mean, 2.9 x 10(2) copies/microl), but the range was great, from (1.2-4.8) x 10(5)/microl in herpetic iridocyclitis. Seventeen percent of cases in which HSV was not suspected to be involved showed a small number of copies of HSV DNA, indicating the unexpected involvement of HSV in these cases.

Conclusions: Real-time PCR is an informative method of diagnosing herpetic eye diseases and evaluating the possible involvement of HSV in other inflammatory ocular diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Aqueous Humor / virology*
  • Child
  • Child, Preschool
  • DNA Primers / chemistry
  • DNA, Viral / analysis
  • Epithelium, Corneal / virology*
  • Female
  • Gene Dosage
  • Genome, Viral
  • Humans
  • Infant
  • Keratitis, Herpetic / diagnosis*
  • Keratitis, Herpetic / virology
  • Male
  • Middle Aged
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Simplexvirus / genetics*
  • Simplexvirus / isolation & purification
  • Tears / virology*


  • DNA Primers
  • DNA, Viral