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通过原位杂交对自然感染猪的肝脏和肝外组织中戊型肝炎病毒进行定位

Localization of swine hepatitis E virus in liver and extrahepatic tissues from naturally infected pigs by in situ hybridization.

作者信息

Choi Changsun, Chae Chanhee

机构信息

Department of Veterinary Pathology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, San 56-1, Shillim-Dong, Kwanak-Gu, 51-742, Seoul, South Korea.

出版信息

J Hepatol. 2003 Jun;38(6):827-32. doi: 10.1016/s0168-8278(03)00115-6.

Abstract

BACKGROUND/AIMS: The objective of this study was to detect and localize the swine hepatitis E virus (HEV) in the liver and extrahepatic tissues from 20 pigs naturally infected with swine HEV.

METHODS

cDNA probe 289 base pairs for swine HEV were generated by reverse transcription-polymerase chain reaction. In situ hybridization with a nonradioactive digoxigenin-labeled cDNA probe was used for the detection of swine HEV in formalin-fixed, paraffin-embedded tissues.

RESULTS

When liver tissues from the pigs naturally infected with swine HEV were hybridized with the nonradioactive digoxigenin-labeled cDNA probe, a strong signal was seen in hepatocytes and bile duct. Positive hybridization signals were also detected in small and large intestine, lymph node, tonsil, spleen, and kidney.

CONCLUSIONS

Swine HEV was detected primarily in the hepatocytes. Swine HEV may also replicate in tissues other than the liver. In situ hybridization described in the present study has greatly facilitated the application of in situ hybridization procedures to the clinical setting, thus allowing for the diagnosis of swine HEV infection while preserving the morphology of the tissue.

摘要

背景/目的:本研究的目的是检测和定位20头自然感染猪戊型肝炎病毒(HEV)的猪肝脏和肝外组织中的病毒。

方法

通过逆转录聚合酶链反应生成猪HEV的289个碱基对的cDNA探针。使用非放射性地高辛标记的cDNA探针对福尔马林固定、石蜡包埋的组织进行原位杂交,以检测猪HEV。

结果

当用非放射性地高辛标记的cDNA探针与自然感染猪HEV的猪肝脏组织杂交时,在肝细胞和胆管中可见强信号。在小肠、大肠、淋巴结、扁桃体、脾脏和肾脏中也检测到阳性杂交信号。

结论

猪HEV主要在肝细胞中检测到。猪HEV也可能在肝脏以外的组织中复制。本研究中描述的原位杂交极大地促进了原位杂交程序在临床环境中的应用,从而在保留组织形态的同时实现猪HEV感染的诊断。

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