Clinical application of real-time polymerase chain reaction for diagnosis of herpetic diseases of the anterior segment of the eye.
OBJECTIVE
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.
Kakimaru-Hasegawa
Akiko
A
Division of Ophthalmology and Visual Science, Faculty of Medicine, Tottori University, Tottori, Japan. kakimaruaki@yahoo.co.jp
Kuo
Chuan-Hui
CH
Komatsu
Naoki
N
Komatsu
Keiko
K
Miyazaki
Dai
D
Inoue
Yoshitsugu
Y
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001082:Aqueous Humor / Q000821:virology*; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis; D019573:Epithelium, Corneal / Q000821:virology*; D005260:Female; D018628:Gene Dosage; D016679:Genome, Viral; D006801:Humans; D007223:Infant; D016849:Keratitis, Herpetic / Q000175:diagnosis* / Q000821:virology; D008297:Male; D008875:Middle Aged; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018139:Simplexvirus / Q000235:genetics* / Q000302:isolation & purification; D013666:Tears / Q000821:virology*
D017931:DNA Primers; D004279:DNA, Viral