Sharma A, Shimeld C
Department of Medical Microbiology, University of Wales College of Medicine, Cardiff.
Br J Ophthalmol. 1997 Sep;81(9):785-8. doi: 10.1136/bjo.81.9.785.
BACKGROUND/AIMS: Herpes simplex virus keratitis (HSK) is the most common cause of corneal blindness in the Western world. Delay in the treatment of HSK can lead to a more significant corneal scar and topical steroid treatment in unsuspected active HSK can lead to corneal melting. Current culture techniques for herpes simplex virus (HSV) take several days and commercially available HSV laboratory based diagnostic techniques such as Herpchek vary in sensitivity. This study was conducted to assess the viability of a new, quicker, and simpler method to diagnose HSK.
Direct immunofluorescence was used in vivo in a masked study to diagnose HSK in mice using a standard slit lamp with cobalt blue illumination. Murine monoclonal fluorescently labelled antibody was applied to the cornea for 10 or 20 minutes and then washed off with phosphate buffered solution. Mice with HSK were stained with either fluorescently labelled monoclonal antibody against HSV or fluorescently labelled monoclonal antibody against cytomegalovirus. Mice with corneal abrasions of non-viral origin were given fluorescently labelled monoclonal antibody against HSV.
Fluorescence was seen only in the mice with HSK given fluorescently labelled monoclonal antibody against HSV. This observation was confirmed upon microscopic immunofluorescent imaging of the corneal epithelial sheets.
In vivo immunofluorescence may be useful in the clinical diagnosis of HSK.
背景/目的：单纯疱疹病毒性角膜炎（HSK）是西方世界角膜盲最常见的病因。HSK治疗延误可导致更严重的角膜瘢痕，而在未怀疑有活动性HSK时进行局部类固醇治疗可导致角膜溶解。目前单纯疱疹病毒（HSV）的培养技术需要数天时间，而基于实验室的市售HSV诊断技术（如Herpchek）在敏感性上存在差异。本研究旨在评估一种新的、更快且更简单的HSK诊断方法的可行性。
在一项盲法研究中，使用直接免疫荧光法在小鼠体内进行HSK诊断，采用配备钴蓝光照明的标准裂隙灯。将鼠单克隆荧光标记抗体应用于角膜10或20分钟，然后用磷酸盐缓冲溶液冲洗掉。患有HSK的小鼠用抗HSV荧光标记单克隆抗体或抗巨细胞病毒荧光标记单克隆抗体染色。对非病毒源性角膜擦伤的小鼠给予抗HSV荧光标记单克隆抗体。
仅在给予抗HSV荧光标记单克隆抗体的患有HSK的小鼠中观察到荧光。对角膜上皮片进行显微镜免疫荧光成像证实了这一观察结果。
体内免疫荧光可能有助于HSK的临床诊断。
