Schmidbauer M, Budka H, Ambros P
Neurological Institute, University of Vienna, Wien.
Virchows Arch A Pathol Anat Histopathol. 1988;414(1):39-43. doi: 10.1007/BF00749736.
Formol-fixed and paraffin-embedded brain tissue of 33 cases of human necrotizing encephalitis was investigated for Herpes simplex virus (HSV) by immunocytochemistry with a polyclonal antiserum, and by in situ hybridization (ISH) with a biotinylated cDNA probe. HSV antigens (VA) were found in various types of cells in the cytoplasm, cellular processes and nuclei. Labelling by ISH was mostly restricted to nuclei and intranuclear inclusions but otherwise matched the distribution of VA. Eighteen of 25 acute cases had HSV antigen detectable by immunocytochemistry, and 18 of the acute cases contained HSV DNA detectable by our ISH technique. However, results differed somewhat between the techniques: three brains negative for VA showed hybridization, and other 3 VA-positive cases remained negative by ISH. Thus 21 brains with acute necrotizing encephalitis were labelled with one or both techniques. In 8 cases with a subacute course (duration of disease was longer than 1 month), HSV antigens were never detectable although 4 brains showed hybridization. All brains labelled by one or both techniques contained nuclear inclusions bodies. Only one case, of subacute course, with inclusion bodies remained unlabelled. Brain tissue of 11 controls, including cytomegalic inclusion body disease, was never labelled. These results demonstrate that immunocytochemistry and ISH are techniques of comparable sensitivity (72%) for detection of HSV in paraffin sections of acute necrotizing encephalitis brains; their combined use enhanced sensitivity, in our hands, to 84%. In cases with a disease course longer than one month, ISH seems to be the method of choice to demonstrate HSV in situ.
采用多克隆抗血清免疫细胞化学法及生物素化cDNA探针原位杂交法（ISH），对33例人类坏死性脑炎的甲醛固定石蜡包埋脑组织进行单纯疱疹病毒（HSV）检测。在各类细胞的细胞质、细胞突起及细胞核中发现了HSV抗原（VA）。ISH标记大多局限于细胞核及核内包涵体，但在其他方面与VA的分布相符。25例急性病例中，18例通过免疫细胞化学可检测到HSV抗原，18例急性病例通过我们的ISH技术可检测到HSV DNA。然而，两种技术的结果略有不同：3例VA阴性的脑显示杂交阳性，另外3例VA阳性病例ISH检测仍为阴性。因此，21例急性坏死性脑炎的脑通过一种或两种技术检测呈阳性。在8例亚急性病程（病程超过1个月）的病例中，尽管4例脑显示杂交阳性，但从未检测到HSV抗原。所有通过一种或两种技术检测呈阳性的脑均含有核内包涵体。仅1例亚急性病程且有包涵体的病例检测呈阴性。11例对照脑组织，包括巨细胞包涵体病，均未检测到阳性。这些结果表明，免疫细胞化学和ISH在检测急性坏死性脑炎脑石蜡切片中的HSV时，敏感性相当（72%）；在我们的研究中，联合使用可将敏感性提高到84%。对于病程超过1个月的病例，ISH似乎是原位检测HSV的首选方法。
