用于检测实验感染的脾切除猪的福尔马林固定石蜡包埋组织中猪血支原体（猪附红细胞体）的原位杂交技术的开发。

Development of in-situ hybridization for the detection of Mycoplasma haemosuis (Eperythrozoon suis) in formalin-fixed, paraffin wax-embedded tissues from experimentally infected splenectomized pigs.

作者信息

Ha S-K, Jung K, Choi C, Ha Y, Song H-C, Lim J-H, Kim S-H, Chae C

机构信息

Departmentof Veterinary Parhology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Seoul.

出版信息

J Comp Pathol. 2005 Nov;133(4):294-7. doi: 10.1016/j.jcpa.2005.06.002. Epub 2005 Oct 5.

DOI:10.1016/j.jcpa.2005.06.002

PMID:16212975

Abstract

Mycoplasma haemosuis DNA was detected in experimentally infected splenectomized pigs by in-situ hybridization (ISH) with a nonradioactive digoxigenin-labelled DNA probe. An 839 base pair DNA probe targeting a 16S rRNA gene was generated by the polymerase chain reaction. Eight 6-week-old pigs were inoculated intraperitoneally with 6 ml of M. haemosuis-infected pig blood and eight negative control pigs were inoculated intraperitoneally with 6 ml of M. haemosuis-free blood. Two pigs from each group were killed for examination at 3, 7, 15 and 30 days post-inoculation (dpi). Red blood cells infected with M. haemosuis were first detected by light microscopy at 3 to 7 dpi. No M. haemosuis was observed in negative control pigs. Hybridization signals were evident in blood from the infected pigs at 3 dpi. The ISH method developed in this study was useful for the detection of M. haemosuis DNA in formalin-fixed, paraffin wax-embedded tissues and may be valuable for studying the pathogenesis of M. haemosuis infection.

摘要

通过使用非放射性地高辛标记的DNA探针进行原位杂交（ISH），在实验感染的脾切除猪中检测到猪血支原体DNA。通过聚合酶链反应产生了一个靶向16S rRNA基因的839碱基对DNA探针。8只6周龄猪经腹腔接种6毫升感染猪血支原体的猪血，8只阴性对照猪经腹腔接种6毫升不含猪血支原体的猪血。每组在接种后3、7、15和30天处死2只猪进行检查。感染猪血支原体的红细胞在接种后3至7天首次通过光学显微镜检测到。阴性对照猪中未观察到猪血支原体。在接种后3天，感染猪的血液中可见杂交信号。本研究开发的ISH方法可用于检测福尔马林固定、石蜡包埋组织中的猪血支原体DNA，并可能对研究猪血支原体感染的发病机制具有重要价值。