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[应用多重聚合酶链反应(PCR)和实时荧光定量PCR检测急性视网膜坏死患者眼内液中的疱疹病毒基因组]

[Detection of herpesvirus genome by multiplex polymerase chain reaction (PCR) and real-time PCR in ocular fluids of patients with acute retinal necrosis].

作者信息

Sugita Sunao, Iwanaga Youichi, Kawaguchi Tatsushi, Futagami Yuri, Horie Shintaro, Usui Tomoka, Yamamoto Sayaka, Sugamoto Yoshiharu, Mochizuki Manabu, Shimizu Norio, Watanabe Ken, Mizukami Miki, Morio Tomohiro

机构信息

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

出版信息

Nippon Ganka Gakkai Zasshi. 2008 Jan;112(1):30-8.

Abstract

PURPOSE

The human herpesvirus (HHV) family consists of types 1 to 8 (HHV1-8). The purpose of this study was to investigate the detection of HHV DNA, especially HSV1 (herpes simplex virus 1, HHV1), HSV2 (herpes simplex virus 2, HHV2), and VZV (varicella-zoster virus, HHV3) in ocular fluids of patients with acute retinal necrosis(ARN).

METHODS

The intraocular genome for HHV1-8 was determined in 19 ocular fluid samples (12 vitreous fluid and 7 aqueous humor samples) taken from ARN patients (n=14). The samples were tested for the presence of virus DNA by two systems of polymerase chain reaction (PCR): the multiplex PCR screening test and real-time quantitative PCR.

RESULTS

Multiplex PCR demonstrated VZV (n=16, 84%), HSV1 (n = 1.5%) or HSV2 (n = 2.11%)genomic DNA in all the samples. In real-time PCR, a high copy number of virus DNA was detected. The virus DNA-positive samples contained Epstein-Barr virus (EBV, HHV4) DNA in 9 of 19 samples (47%). No HHV6-8 DNA was detected in the ocular samples, and no virus DNA was detected in the serum samples.

CONCLUSIONS

The genome for HHV1-3 was detected in the patients with ARN. All cases contained a high copy number for the virus DNA that indicates viral replication. PCR systems are useful for determing whether virus infections are associated with uveitis.

摘要

目的

人类疱疹病毒(HHV)家族由1型至8型(HHV1 - 8)组成。本研究的目的是调查急性视网膜坏死(ARN)患者眼内液中HHV DNA的检测情况,尤其是单纯疱疹病毒1型(HSV1,HHV1)、单纯疱疹病毒2型(HSV2,HHV2)和水痘 - 带状疱疹病毒(VZV,HHV3)。

方法

对14例ARN患者的19份眼内液样本(12份玻璃体液和7份房水样本)进行HHV1 - 8的眼内基因组检测。通过两种聚合酶链反应(PCR)系统检测样本中病毒DNA的存在:多重PCR筛选试验和实时定量PCR。

结果

多重PCR在所有样本中均检测到VZV(n = 16,84%)、HSV1(n = 1.5%)或HSV2(n = 2.

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