Brown C C, Meyer R F, Olander H J, House C, Mebus C A
Foreign Animal Disease Diagnostic Laboratory, National Veterinary Services Laboratories, APHIS-USDA, Greenport, New York 11944.
Can J Vet Res. 1992 Jul;56(3):189-93.
Eight calves were exposed in an aerosol chamber to nebulized foot-and-mouth disease virus. Two control animals were exposed in a similar manner to cell culture media only. Animals were euthanized at intervals and various tissues examined by in situ hybridization using a biotinylated RNA probe corresponding to a portion of the viral gene coding for the polymerase enzyme. By this technique large amounts of viral nucleic acid were found in coronary band, interdigital cleft and tongue as early as six hours postexposure, indicating a very rapid delivery from the portal of entry to the predilection sites for lesion development. This occurred well before the onset of viremia which by virus isolation was not detectable until 30 hours postexposure. The in situ hybridization signal in these tissues decreased in intensity and extent with time to focally positive areas, occasionally surrounding a vesicle. Other epidermal sites not normally thought of as sites for foot-and-mouth lesion development, such as carpus and eyelid, also had some viral nucleic acid detectable at various time intervals. In the lung by in situ hybridization, alveolar septa had viral nucleic acid early in infection (6-18 h postexposure) while later (36-96 h postexposure), the in situ hybridization signal was prominent in alveolar macrophages.
八头小牛在气溶胶室内暴露于雾化的口蹄疫病毒。两只对照动物以类似方式仅暴露于细胞培养基。在不同时间间隔对动物实施安乐死,并使用与编码聚合酶的病毒基因部分相对应的生物素化RNA探针,通过原位杂交检查各种组织。通过该技术,早在暴露后6小时就在冠状带、趾间裂和舌中发现了大量病毒核酸,这表明从进入门户到病变发展的偏好部位的传递非常迅速。这发生在病毒血症发作之前,直到暴露后30小时通过病毒分离才检测到病毒血症。这些组织中的原位杂交信号强度和范围随时间下降,变为局灶性阳性区域,偶尔围绕一个水疱。其他通常不被认为是口蹄疫病变发展部位的表皮部位,如腕关节和眼睑,在不同时间间隔也可检测到一些病毒核酸。通过原位杂交在肺中发现,感染早期(暴露后6-18小时)肺泡隔中有病毒核酸,而后期(暴露后36-96小时),原位杂交信号在肺泡巨噬细胞中很明显。