In situ hybridizaiton in Viral hepatitis

Abstract

In situ hybridizaiton (ISH) is a sensitive and specific technique for detecting nucleic acids in intact cells. Visualization of the target sequences by autoradiography or immunohistochemistry allows their precise subcellular localization and quantitation. The application of ISH techniques has contributed to the understanding of the complex replicative cycle of hepatitis B virus. More recently, hepatitis delta and C virus replication has also been studied by this technique. ISH‐based assays have finally been used to follow the replication of cytomegalovirus within the transplanted liver. Although ISH is a powerful tool for the molecular biologist, its clinical significance in the diagnosis and prognosis of human I hepatitis virus infections has yet to be fully evaluated.

Imaging of Hepatitis B Virus Nucleic Acids: Current Advances and Challenges

“…Later, biotin-labelled probes were used together with an avidine–peroxidase complex [ 37 , 38 ], as well as digoxigenin-labelled probes applied with alkaline phosphatase-conjugated antibodies directed against digoxigenin [ 39 ]. The studies identified the cytoplasm as the site where most HBV DNA localise and exhibited two distinct patterns, either diffused staining in the majority of hepatocytes or staining confined to foci [ 69 ]. By treating liver sections with RNase and using ISH without denaturation, probes which target the single-stranded region of rcDNA predominantly showed staining in the cytoplasm [ 35 ].…”