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定量聚合酶链反应检测葡萄膜炎患者眼内液中爱泼斯坦-巴尔病毒的基因组DNA。

Quantitative PCR for the detection of genomic DNA of Epstein-Barr virus in ocular fluids of patients with uveitis.

作者信息

Yamamoto Sayaka, Sugita Sunao, Sugamoto Yoshiharu, Shimizu Norio, Morio Tomohiro, Mochizuki Manabu

机构信息

Department of Ophthalmology & Visual Science, Tokyo Medical and Dental University, Tokyo, Japan.

Department of Virology, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.

出版信息

Jpn J Ophthalmol. 2008 Nov-Dec;52(6):463-467. doi: 10.1007/s10384-008-0589-8. Epub 2008 Dec 17.

Abstract

PURPOSE

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.

摘要

目的

检测眼内液中爱泼斯坦-巴尔病毒(EBV)的基因组DNA,并分析EBV在葡萄膜炎中的临床相关性。

方法

在获得知情同意后,从55例葡萄膜炎患者中采集眼内液(30份房水和30份玻璃体样本)。使用定性多重聚合酶链反应(PCR)和定量实时PCR检测样本中的EBV DNA。使用补体结合试验检测EBV抗体。

结果

使用多重PCR在60份样本中的17份(28%)以及55例患者中的16例(29%)检测到EBV DNA。然而,在这17份样本中,只有3份通过实时PCR显示出EBV DNA的拷贝数显著较高。所有患者的血清中均未检测到EBV DNA。所有患者血清中的EBV特异性抗体均为阳性,但玻璃体中未检测到。仅在玻璃体样本中显示出EBV基因组DNA的患者中,玻璃体抗EBV抗体呈阳性。

结论

通过定性PCR在许多葡萄膜炎患者的眼内液中检测到EBV DNA,但只有一小部分患者在定量实时PCR中显示出高病毒载量,表明该病毒仅在少数患者中复制。

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