一种用于检测肝组织中 HBV RNA、DNA 和 cccDNA 的显色原位杂交 (CISH) 检测方法。
A Chromogenic In Situ Hybridization (CISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Liver Tissue.
机构信息
Centre for Research in Therapeutic Solutions, Biomedical Sciences, Faculty of Science and Technology, University of Canberra, Canberra, ACT, Australia.
Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
出版信息
Methods Mol Biol. 2024;2837:137-148. doi: 10.1007/978-1-0716-4027-2_12.
Hepatitis B virus (HBV) developed highly intricates mechanisms exploiting host resources for its multiplication within a constrained genetic coding capacity. With the aid of a series of classical analytical methods such as ultrafiltration, and Southern and Northern blots, a general framework of HBV life cycle has been established. However, this picture still lacks many key histological contexts which involves pathophysiological changes of hepatocytes, non-parenchymal cells, infiltrated leukocytes, and associated extracellular matrix. Here, we describe a CISH protocol modified from the ViewRNA assay that allows direct visualization of HBV RNA, DNA, and cccDNA in liver tissue of chronic hepatitis B patients. By coupling it with immunohistochemistry and other histological stains, much richer information regarding the HBV-induced pathological changes can be harvested.
乙型肝炎病毒 (HBV) 开发了高度复杂的机制，利用宿主资源在有限的遗传编码能力内进行繁殖。借助一系列经典的分析方法，如超滤、Southern 和 Northern 印迹，HBV 生命周期的一般框架已经建立。然而，这一图景仍然缺乏许多关键的组织学背景，这些背景涉及肝细胞、非实质细胞、浸润的白细胞以及相关细胞外基质的病理生理变化。在这里，我们描述了一种从 ViewRNA 检测方法修改而来的 CISH 方案，该方案允许直接观察慢性乙型肝炎患者肝组织中的 HBV RNA、DNA 和 cccDNA。通过将其与免疫组织化学和其他组织学染色相结合，可以获得更多关于 HBV 诱导的病理变化的信息。
Zotero 插件