Real-time polymerase chain reaction for the diagnosis of necrotizing herpes stromal keratitis.
OBJECTIVE
To design, optimize and validate a rapid, internally controlled real-time polymerase chain reaction (RT-PCR) test for herpes simplex virus (HSV) in the diagnosis of necrotizing herpes stromal keratitis.


METHODS
Tears alone or together with corneal epithelium scrapings from 30 patients (30 eyes) suspected of necrotizing herpes stromal keratitis were tested for HSV DNA by RT-PCR. The samples were collected during the first visit and then on the subsequent 7, 14, 28, 42, and 56d. The symptoms of the patients were scored before treatment to determine the correlation between HSV concentration in the corneal epithelium scrapings and clinical scores.


RESULTS
The positive rate (46.4%) in the corneal epithelium group before the therapy was significantly higher than that (13.3%) in the tears group (P=0.006). There were 13 positive HSV patients before the therapy, the concentration of HSV DNA in corneal epithelium scrapings group was significantly higher than that in the tears group (paired t-test, P=0.0397). Multilevel mixed-effects model analysis showed that the difference between the corneal epithelium scrapings group and the tears group was statistically significant (P=0.0049). The Spearman rank correlation analysis indicated a positive correlation between the HSV concentration in the corneal epithelium scrapings and clinical scores before the treatment (r=0.844, P<0.0001).


CONCLUSIONS
RT-PCR appears to be a powerful molecular tool for the diagnosis of necrotizing herpes stromal keratitis.
Department of Ophthalmology, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, Jiangsu Province, China.
Jun-Xin
JX
Ma
Department of Ophthalmology, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, Jiangsu Province, China.
Lin-Nong
LN
Wang
Department of Ophthalmology, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, Jiangsu Province, China.
Ru-Xia
RX
Zhou
Department of Clinical Nuclear Medicine Center, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, Jiangsu Province, China.
Yang
Y
Yu
Department of Clinical Nuclear Medicine Center, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, Jiangsu Province, China.
Tong-Xin
TX
Du
corneal epithelium scrapings; necrotizing herpes stromal keratitis; real-time polymerase chain reaction; tears