Quantitative PCR for the detection of genomic DNA of Epstein-Barr virus in ocular fluids of patients with uveitis.
OBJECTIVE
To measure the genomic DNA from Epstein-Barr virus (EBV) in ocular fluids and to analyze the clinical relevance of EBV in uveitis.


METHODS
Intraocular fluids (30 aqueous humor and 30 vitreous fluid samples) were taken from 55 patients with uveitis after informed consent was obtained. Samples were assayed for EBV DNA using qualitative multiplex polymerase chain reaction (PCR) and quantitative real-time PCR. Antibodies to EBV were examined using a complement fixation test.


RESULTS
EBV DNA was detected in 17 of 60 samples (28%) and 16 of 55 patients (29%) using multiplex PCR. However, only three of the 17 samples showed significantly high copy numbers of EBV DNA with real-time PCR. EBV DNA was not detected in the serum of all patients. EBV-specific antibodies were positive in the serum of all patients, but not in the vitreous fluid. Vitreous anti-EBV antibodies were positive only in patients displaying genomic DNA of EBV in the vitreous samples.


CONCLUSIONS
EBV DNA was detected by qualitative PCR in ocular fluids of many uveitis patients, but only a small proportion of patients showed high viral loads on quantitative real-time PCR, indicating that replication of the virus takes place only in a few patients.
Yamamoto
Sayaka
S
Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University, Tokyo, Japan.
Sugita
Sunao
S
Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University, Tokyo, Japan.
Sugamoto
Yoshiharu
Y
Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University, Tokyo, Japan.
Shimizu
Norio
N
Department of Virology, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.
Morio
Tomohiro
T
Center for Cell Therapy, Tokyo Medical and Dental University, Tokyo, Japan.
Mochizuki
Manabu
M
Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University, Tokyo, Japan. m.manabu.oph@tmd.ac.jp.
D000368:Aged; D000914:Antibodies, Viral / Q000032:analysis / Q000097:blood; D001082:Aqueous Humor / Q000821:virology*; D003199:Computer Systems; D004279:DNA, Viral / Q000032:analysis*; D018628:Gene Dosage; D016679:Genome, Viral; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000276:immunology; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D014605:Uveitis / Q000821:virology*; D019562:Viral Load; D014822:Vitreous Body / Q000821:virology*
D000914:Antibodies, Viral; D004279:DNA, Viral