Guedes Roberto M C, Gebhart Connie J, Winkelman Nathan L, Mackie-Nuss Rebecca A C, Marsteller Thomas A, Deen John
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Minnesota, Saint Paul 55108, USA.
Can J Vet Res. 2002 Apr;66(2):99-107.
The objectives of this study were: (1) to compare 2 methods of serology; (2) to compare 3 histologic techniques; and (3) to compare 2 methods of detecting shedding in pigs experimentally challenged with Lawsonia intracellularis. The sensitivities of these tests were determined by the detection of infection. Forty 5-week-old pigs were inoculated on day 0 with intestinal homogenate from pigs with proliferative enteropathy (PE). Clinical evaluation was done on day 7 and daily from day 14 to 28 postinoculation. Fecal shedding of L. intracellularis was monitored by use of polymerase chain reaction (PCR) analysis and immunoperoxidase staining at 7-day intervals. Serum was obtained on days 0 and 28 for serologic testing by glass slide and tissue culture indirect fluorescent antibody tests. At euthanasia on day 28, gross intestinal lesions were evaluated and ileum samples collected for histologic analyses. Ileal histologic sections from each animal were stained by hematoxylin and eosin, Warthin-Starry silver stain, and immunohistochemistry (IHC). Of the 40 pigs, 36 had gross lesions typical of PE at necropsy. The percentage of agreement between the 2 serologic methods was 94.4%. Immunoperoxidase stain of fecal smears was more sensitive than PCR for detecting fecal shedding, especially on day 21 (89.5% and 60.5%, respectively) and day 28 (59.4% and 37.5%, respectively) post-inoculation. The IHC stain was much more sensitive for detecting infection than the routinely used hematoxylin and eosin and Warthin-Starry silver stains. In conclusion, in experimentally infected pigs, both serologic methods were appropriate techniques for detecting infection. For fecal samples, PCR has low sensitivity. Immunohistochemistry is the best diagnostic tool for formalin-fixed samples.
(1)比较两种血清学方法;(2)比较三种组织学技术;(3)比较两种检测经细胞内劳森菌实验性攻毒的猪排毒情况的方法。这些检测的敏感性通过感染检测来确定。40头5周龄仔猪于第0天接种来自患有增生性肠炎(PE)的猪的肠道匀浆。在第7天进行临床评估,并在接种后第14天至28天每天进行评估。每隔7天使用聚合酶链反应(PCR)分析和免疫过氧化物酶染色监测细胞内劳森菌的粪便排毒情况。在第0天和第28天采集血清,通过玻片和组织培养间接荧光抗体试验进行血清学检测。在第28天安乐死时,评估肠道大体病变并采集回肠样本进行组织学分析。每只动物的回肠组织学切片用苏木精和伊红、Warthin-Starry银染色和免疫组织化学(IHC)染色。40头猪中,36头在尸检时有典型的PE大体病变。两种血清学方法之间的一致性百分比为94.4%。粪便涂片的免疫过氧化物酶染色在检测粪便排毒方面比PCR更敏感,尤其是在接种后第21天(分别为89.5%和60.5%)和第28天(分别为59.4%和37.5%)。IHC染色在检测感染方面比常规使用的苏木精和伊红以及Warthin-Starry银染色敏感得多。总之,在实验感染的猪中,两种血清学方法都是检测感染的合适技术。对于粪便样本,PCR敏感性较低。免疫组织化学是福尔马林固定样本的最佳诊断工具。