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用于检测和定位猪沙眼衣原体的原位杂交技术。

In situ hybridization for the detection and localization of swine Chlamydia trachomatis.

作者信息

Chae C, Cheon D S, Kwon D, Kim O, Kim B, Suh J, Rogers D G, Everett K D, Andersen A A

机构信息

Department of Veterinary Pathology, College of Veterinary Medicine, Seoul National University, Suwon, Republic of Korea.

出版信息

Vet Pathol. 1999 Mar;36(2):133-7. doi: 10.1354/vp.36-2-133.

Abstract

Gnotobiotic piglets were inoculated intralaryngeally with swine Chlamydia trachomatis strain R33 or orally with swine C. trachmatis strain R27. Archived formalin-fixed, paraffin-embedded tissues from piglets euthanatized 4-7 days postinoculation were examined by in situ hybridization for C. trachomatis nucleic acid using a nonradioactive digoxigenin-labeled DNA probes that targeted specific ribosomal RNA or omp1 mRNA molecules of the swine C. trachomatis strains. Positive hybridization signals were detected in bronchial epithelial cells, bronchiolar epithelial cells, pneumocytes, alveolar and interstitial macrophages, and jejunal and ileal enterocytes. Chlamydia-infected cells had a strong signal that was confined to the intracytoplasmic inclusions. Positive hybridization signals were not detected in tissue sections from an uninfected control piglet or in C. psittaci-infected sheep placenta. The morphology of host cells was preserved despite the relatively high temperature required in parts of the incubation procedure. The data indicate that in situ hybridization can be used to detect swine C. trachomatis in formalin-fixed, paraffin-embedded tissue specimens.

摘要

无菌仔猪经喉内接种猪沙眼衣原体R33株,或经口接种猪沙眼衣原体R27株。对接种后4 - 7天安乐死的仔猪的存档福尔马林固定、石蜡包埋组织,使用针对猪沙眼衣原体菌株特异性核糖体RNA或omp1 mRNA分子的非放射性地高辛标记DNA探针,通过原位杂交检测沙眼衣原体核酸。在支气管上皮细胞、细支气管上皮细胞、肺细胞、肺泡和间质巨噬细胞以及空肠和回肠肠细胞中检测到阳性杂交信号。衣原体感染的细胞有强烈信号,局限于胞质内包涵体。在未感染对照仔猪的组织切片或鹦鹉热衣原体感染的绵羊胎盘中未检测到阳性杂交信号。尽管孵育过程部分需要相对较高的温度,但宿主细胞的形态得以保留。数据表明,原位杂交可用于检测福尔马林固定、石蜡包埋组织标本中的猪沙眼衣原体。

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