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使用生物素化DNA探针通过DNA-DNA杂交技术检测疱疹性眼病中的1型单纯疱疹病毒。

Detection of herpes simplex virus type 1 in herpetic ocular diseases by DNA-DNA hybridization using a biotinylated DNA probe.

作者信息

Nago R, Hayashi K, Ochiai H, Kubota Y, Niwayama S

机构信息

Department of Ophthalmology, Toyama Medical and Pharmaceutical University, Japan.

出版信息

J Med Virol. 1988 Jul;25(3):259-70. doi: 10.1002/jmv.1890250303.

Abstract

A diagnostic hybridization assay for detecting herpes simplex virus type 1 (HSV-1) in ocular specimens was developed using cloned viral DNA as a probe. This hybridization assay is based on visualizing a biotinylated probe that is hybridized to the target DNA by a streptavidin/alkaline phosphatase system. The time required for performing this assay system is only two days. This assay system could detect a probe which had been hybridized to as little as 1 pg of homologous DNA and did not cross-react with DNA of other human herpes viruses except that of herpes simplex virus type 2 (HSV-2) which showed weak cross-reactivity. The assay system was applied to experimental keratitis in albino rabbits and clinical specimens. In experimental keratitis in rabbits it was possible to detect HSV-1 DNA in the eye swab samples at least until the ninth day after virus inoculation. Five clinical specimens collected from patients with corneal ulcer or blepharitis contained HSV-1 DNA in spite of the failure of demonstration of viral antigen and/or virus isolation in two cases.

摘要

利用克隆的病毒DNA作为探针,开发了一种用于检测眼标本中单纯疱疹病毒1型(HSV-1)的诊断杂交试验。这种杂交试验基于通过链霉亲和素/碱性磷酸酶系统可视化与靶DNA杂交的生物素化探针。进行该试验系统所需的时间仅为两天。该试验系统能够检测到与低至1 pg同源DNA杂交的探针,并且除了单纯疱疹病毒2型(HSV-2)显示出弱交叉反应外,不与其他人类疱疹病毒的DNA发生交叉反应。该试验系统应用于白化兔的实验性角膜炎和临床标本。在兔的实验性角膜炎中,至少在病毒接种后第九天能够在眼拭子样本中检测到HSV-1 DNA。从角膜溃疡或睑缘炎患者收集的五份临床标本中含有HSV-1 DNA,尽管在两例病例中未能证明病毒抗原和/或病毒分离。

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