Use of multiplex PCR and real-time PCR to detect human herpes virus genome in ocular fluids of patients with uveitis.
OBJECTIVE
To measure the genomic DNA of human herpes viruses (HHV) in the ocular fluids and to analyse the clinical relevance of HHV in uveitis.


METHODS
After informed consent was obtained, a total of 111 ocular fluid samples (68 aqueous humour and 43 vitreous fluid samples) were collected from 100 patients with uveitis. The samples were assayed for HHV-DNA (HHV1-8) by using two different polymerase chain reaction (PCR) assays, qualitative PCR (multiplex PCR) and quantitative PCR (real-time PCR).


RESULTS
In all of the patients with acute retinal necrosis (n = 16) that were tested, either the HSV1 (n = 2), HSV2 (n = 3), or VZV (n = 11) genome was detected. In all patients, high copy numbers of the viral DNA were also noted, indicating the presence of viral replication. In another 10 patients with anterior uveitis with iris atrophy, the VZV genome was detected. When using multiplex PCR, EBV-DNA was detected in 19 of 111 samples (17%). However, real-time PCR analysis of EBV-DNA indicated that there were only six of the 19 samples that had significantly high copy numbers. The cytomegalovirus (CMV) genome was detected in three patients with anterior uveitis of immunocompetent patients and in one immunocompromised CMV retinitis patient. In addition, one patient with severe unilateral panuveitis had a high copy number of HHV6-DNA. There was no HHV7- or HHV8-DNA detected in any of the samples.


CONCLUSIONS
A qualitative multiplex PCR is useful in the screening of viral infections. However, the clinical relevance of the virus infection needs to be evaluated by quantitative real-time PCR.
Sugita
S
S
Department of Ophthalmology & Visual Science, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.
Shimizu
N
N
Watanabe
K
K
Mizukami
M
M
Morio
T
T
Sugamoto
Y
Y
Mochizuki
M
M
D018138:Alphaherpesvirinae / Q000235:genetics / Q000302:isolation & purification*; D001082:Aqueous Humor / Q000821:virology; D004279:DNA, Viral / Q000032:analysis*; D015828:Eye Infections, Viral / Q000175:diagnosis* / Q000821:virology; D016679:Genome, Viral; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D014605:Uveitis / Q000821:virology*; D014822:Vitreous Body / Q000821:virology
D004279:DNA, Viral