Huntley J F J, Whitlock R H, Bannantine J P, Stabel J R
Department of Veterinary Pathology, Iowa State University, Ames, IA, USA.
Vet Pathol. 2005 Jan;42(1):42-51. doi: 10.1354/vp.42-1-42.
Tissues and fecal material were collected from 14 North American bison (Bison bison) that were suspected of having Johne's disease and analyzed for the presence of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis). Sections of ileum, ileal-cecal lymph node, and three sequential sections of jejunum with their associated mesenteric lymph nodes were taken from each animal. Fecal culture indicated that 5 of 14 (35.7%) animals were infected, whereas cultures from tissues detected 12 of 14 (85.7%) animals as infected and 59 of 111 (53.2%) of the tissues as positive for M. paratuberculosis. Polymerase chain reaction analysis identified infection in 14 of 14 (100%) animals and in 91 of 112 (81.2%) tissues. In addition, tissues were processed for Ziehl-Neelsen acid-fast staining, auramine O/acridine orange fluorescent staining, and immunohistochemical staining. Ziehl-Neelsen and auramine O staining identified 7 of 14 (50%) and 5 of 14 (35.7%) animals as infected and 24 of 112 (21.4%) and 28 of 112 (25%) tissues as positive, respectively. Immunohistochemical analyses of bison tissues, using antisera collected from rabbits immunized with four different preparations of M. paratuberculosis, identified a greater percentage of infected animals (ranging from 57 to 93%) and positive tissues (ranging from 28 to 46%). Collectively, these data indicate that DNA-based detection of M. paratuberculosis was more sensitive than bacterial culture or staining, identified infection in all the bison, and detected the greatest number of positive tissues within each animal.
从14头疑似患有副结核的北美野牛（美洲野牛）身上采集组织和粪便样本，分析其中副结核分枝杆菌（副结核菌）的存在情况。从每头动物身上获取回肠、回盲淋巴结以及三段连续的空肠及其相关肠系膜淋巴结的组织切片。粪便培养表明，14头动物中有5头（35.7%）被感染，而组织培养检测出14头动物中有12头（85.7%）被感染，111份组织中有59份（53.2%）对副结核菌呈阳性反应。聚合酶链反应分析确定14头动物中有14头（100%）被感染，112份组织中有91份（81.2%）被感染。此外，对组织进行齐-尼抗酸染色、金胺O/吖啶橙荧光染色和免疫组织化学染色处理。齐-尼染色和金胺O染色分别确定14头动物中有7头（50%）和5头（35.7%）被感染，112份组织中有24份（21.4%）和28份（25%）呈阳性。使用从用四种不同副结核菌制剂免疫的兔子身上采集的抗血清对野牛组织进行免疫组织化学分析，结果显示被感染动物的比例更高（57%至93%），阳性组织的比例也更高（28%至46%）。总体而言，这些数据表明基于DNA的副结核菌检测比细菌培养或染色更敏感，能确定所有野牛均被感染，且能检测出每头动物中阳性组织的数量最多。
