Detection of Aujeszky's disease virus in paraffin-embedded tissues by in situ hybridization using non-radionctive probes

摘要

The Aujeszky's disease (AD), also called pseudorabies, is primarily an infectious disease of swine which is caused by a suid herpesvirus 1 belonging to the subfamily Alphaherpesvirinae, family Herpesvirdae. It was first described in the United States in 1813 in cattle and the etiological agent was firstly recognised as a virus by Aujeszky in Hungary in 1902. Now the disease occurres worldwide. In Korea it was first identified in 1987 and the disease is now more prevalent than before. Laboratory diagnosis of AD can be achieved using serology, histopathology, rabbit inoculation, fluorescent antibody test, immunohistochemistry, polymerase chain reaction(PCR), in situ hybridisation(ISH) and virus isolation. The use of immunohistochemical methods for the detection of viral antigen in formalin-fixed and paraffin-embedded tissues has advantages such as direct visualisation and localisation of virus simultaneously with the evaluation of hislological lesions as well as possibility of retrospective diagnosis As with immunohistochemistry, in situ hybridisation allows the detection and localisation of virus within cells and tissues, but a specific genomic probe instead of the antibody against viral antigen is used to detect specific viral nucleic acids. Recently, a non-radioactive in situ hybridisation method. using a digoxigenin(DIG)-labeled cDNA probe, was reported for the detection of specific antigen in formalin-fixed paraffin-embedded tissue. The aim of the present study was to make DIG-labeled DNA probes with a specificity to AD viral genome and to use the probes for the detection of viral nucleic acids in formalin-fixed and paraffin-embedded tissue sections from pigs naturally infected with Aujeszky's disease virus.