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猪增生性肠炎细胞内微生物的特异性原位杂交

Specific in situ hybridization of the intracellular organism of porcine proliferative enteropathy.

作者信息

Gebhart C J, McOrist S, Lawson G H, Collins J E, Ward G E

机构信息

Department of Veterinary PathoBiology, University of Minnesota, St. Paul.

出版信息

Vet Pathol. 1994 Jul;31(4):462-7. doi: 10.1177/030098589403100409.

Abstract

The identity of the intracellular bacteria found in the enterocytes of pigs with proliferative enteropathy was investigated using specific DNA probes to various Campylobacter species and to a novel organism, ileal symbiont intracellularis. The ilea from pigs (Nos. 1-7) that were diagnosed by routine histopathology as having proliferative enteropathy were used. Diagnosis was made on the basis of proliferation of the enterocytes on hematoxylin and eosin-stained sections and the presence of large numbers of intracellular curved organisms on Warthin-Starry silver-stained sections. Four of these pigs (Nos. 1-4) had the chronic form of the disease, porcine intestinal adenomatosis, and three (Nos. 5-7) had the acute form, proliferative hemorrhagic enteropathy. An additional three normal pigs (Nos. 8-10) were obtained from three separate farms with no history of proliferative enteropathy. Frozen ileal sections were examined by in situ hybridization with DNA probes specific for ileal symbiont intracellularis and the three porcine intestinal Campylobacter species, C. coli, C. hyointestinalis, and C. mucosalis. In all seven pigs with either the intestinal adenomatosis or hemorrhagic enteropathy form of the disease, a DNA probe specific for ileal symbiont intracellularis hybridized to localized foci in the apical cytoplasm of ileal enterocytes. These hybridization sites corresponded to the location of intracellular bacteria in silver-stained sections of adjacent tissue. Sections from the three normal pigs tested with this probe and from all pigs tested with the Campylobacter species-specific DNA probes showed no specific hybridization reactions. The identity of the intracellular organism in these diseased pigs is ileal symbiont intracellularis.

摘要

利用针对各种弯曲杆菌属物种以及一种新型生物体——回肠细胞内共生菌的特异性DNA探针,对患有增生性肠炎的猪的肠上皮细胞中发现的细胞内细菌进行了鉴定。使用了经常规组织病理学诊断为患有增生性肠炎的猪(编号1 - 7)的回肠。诊断依据是苏木精和伊红染色切片中肠上皮细胞的增生,以及Warthin - Starry银染色切片中大量细胞内弯曲生物体的存在。其中四只猪(编号1 - 4)患有慢性形式的疾病——猪肠道腺瘤病,三只猪(编号5 - 7)患有急性形式的疾病——增生性出血性肠炎。另外从三个没有增生性肠炎病史的不同农场获得了三只正常猪(编号8 - 10)。通过与针对回肠细胞内共生菌以及三种猪肠道弯曲杆菌物种——大肠弯曲杆菌、猪肠弯曲杆菌和黏膜弯曲杆菌的DNA探针进行原位杂交,对冷冻的回肠切片进行了检查。在所有七只患有肠道腺瘤病或出血性肠炎形式疾病的猪中,一种针对回肠细胞内共生菌的DNA探针与回肠肠上皮细胞顶端细胞质中的局部病灶杂交。这些杂交位点与相邻组织银染色切片中细胞内细菌的位置相对应。用该探针检测的三只正常猪的切片以及用弯曲杆菌属物种特异性DNA探针检测的所有猪的切片均未显示出特异性杂交反应。这些患病猪中细胞内生物体的身份是回肠细胞内共生菌。

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