Inoue Satoshi, Sato Yuko, Hasegawa Hideki, Noguchi Akira, Yamada Akio, Kurata Takeshi, Iwasaki Takuya
Department of Veterinary Science, National Institute of Infectious Diseases, Tokyo, Japan.
Pathol Int. 2003 Aug;53(8):525-33. doi: 10.1046/j.1440-1827.2003.01511.x.
Diagnosis of rabies is routinely confirmed by detection of rabies virus antigens in acetone-fixed frozen brain tissues or imprint smears using an immunofluorescence method with commercial antirabies virus antibodies. Since recent molecular analyses disclosed wide heterogeneity in the genome sequences of rabies virus strains and related lyssaviruses, it is necessary to confirm the presence of common epitopes in these lyssaviruses. In this study we confirmed the presence of cross-reactive antigens of various lyssaviruses in paraffin sections of formalin-fixed tissue using a monospecific rabbit antiserum prepared by immunization with a recombinant nucleoprotein of rabies virus. By immunohistochemical application, the antigen was detected predominantly in the cytoplasm of neurons in the brains of mice infected with rabies virus, Duvenhage virus, Mokola virus and European bat lyssavirus-1, while no cross-reaction was observed in uninfected humans and animals including dogs, bats, and raccoons. In addition, we examined one autopsy case that was infected in a rabies-endemic nation and developed the clinical manifestation of rabies after returning to Japan in 1970, and found that the antigen was well preserved in paraffin sections of formalin-fixed tissues. Thus, this suggests that the lyssavirus-specific antigen is recognized by the monospecific antibody against rabies virus nucleoprotein, and that this cross-reactive antigen is detectable on formalin-fixed paraffin-embedded tissues by immunohistochemical analysis.
狂犬病的诊断通常是通过使用商业抗狂犬病病毒抗体的免疫荧光法，在丙酮固定的冷冻脑组织或印片涂片中检测狂犬病病毒抗原来确诊。由于最近的分子分析揭示了狂犬病病毒株和相关狂犬病病毒属病毒的基因组序列存在广泛的异质性，因此有必要确认这些狂犬病病毒属病毒中共同表位的存在。在本研究中，我们使用通过用狂犬病病毒重组核蛋白免疫制备的单特异性兔抗血清，在福尔马林固定组织的石蜡切片中证实了各种狂犬病病毒属病毒存在交叉反应性抗原。通过免疫组织化学应用，在感染狂犬病病毒、杜文海格病毒、莫科拉病毒和欧洲蝙蝠狂犬病病毒1型的小鼠大脑神经元细胞质中主要检测到该抗原，而在未感染的人类和动物（包括狗、蝙蝠和浣熊）中未观察到交叉反应。此外，我们检查了一例于1970年在狂犬病流行国家感染并在返回日本后出现狂犬病临床表现的尸检病例，发现该抗原在福尔马林固定组织的石蜡切片中保存良好。因此，这表明狂犬病病毒属特异性抗原可被针对狂犬病病毒核蛋白的单特异性抗体识别，并且这种交叉反应性抗原可通过免疫组织化学分析在福尔马林固定石蜡包埋组织上检测到。
