A Chromogenic In Situ Hybridization (CISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Liver Tissue

Xiaonan Zhang; Wei Lu; Yanling Feng; Zhanqing Zhang; Zhenghong Yuan

doi:10.1007/978-1-0716-4027-2_12

A Chromogenic In Situ Hybridization (CISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Liver Tissue

Xiaonan Zhang
, Wei Lu
, Yanling Feng
, Zhanqing Zhang
, Zhenghong Yuan

Research output: A Conference proceeding or a Chapter in Book › Chapter › peer-review

Abstract

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.

Original language	English
Title of host publication	Hepatitis B Virus
Subtitle of host publication	Methods and Protocols
Editors	Haitao Guo, Andrea Cuconati
Place of Publication	Netherlands
Publisher	Springer
Chapter	12
Pages	137-148
Number of pages	12
Volume	2837
Edition	2
ISBN (Electronic)	9781071640296
ISBN (Print)	9781071640265
DOIs	https://doi.org/10.1007/978-1-0716-4027-2_12
Publication status	Published - 2024

Publication series

Name	Methods in molecular biology (Clifton, N.J.)
Publisher	Humana Press
ISSN (Print)	1064-3745

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

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10.1007/978-1-0716-4027-2_12

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Zhang, X., Lu, W., Feng, Y., Zhang, Z., & Yuan, Z. (2024). A Chromogenic In Situ Hybridization (CISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Liver Tissue. In H. Guo, & A. Cuconati (Eds.), Hepatitis B Virus: Methods and Protocols (2 ed., Vol. 2837, pp. 137-148). (Methods in molecular biology (Clifton, N.J.)). Springer. https://doi.org/10.1007/978-1-0716-4027-2_12

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title = "A Chromogenic In Situ Hybridization (CISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Liver Tissue",

abstract = "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.",

keywords = "cccDNA, CISH, HBV, Immunohistochemistry",

author = "Xiaonan Zhang and Wei Lu and Yanling Feng and Zhanqing Zhang and Zhenghong Yuan",

note = "Publisher Copyright: {\textcopyright} 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature. Funding Information: This work is supported by the Chinese National Natural Science Foundation (81873962, 32070152) and by Australian Centre for HIV and Hepatitis Virology Research (ACH2 ). Publisher Copyright: {\textcopyright} The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024.",

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isbn = "9781071640265",

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Zhang, X, Lu, W, Feng, Y, Zhang, Z & Yuan, Z 2024, A Chromogenic In Situ Hybridization (CISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Liver Tissue. in H Guo & A Cuconati (eds), Hepatitis B Virus: Methods and Protocols. 2 edn, vol. 2837, Methods in molecular biology (Clifton, N.J.), Springer, Netherlands, pp. 137-148. https://doi.org/10.1007/978-1-0716-4027-2_12

A Chromogenic In Situ Hybridization (CISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Liver Tissue. / Zhang, Xiaonan; Lu, Wei; Feng, Yanling et al.
Hepatitis B Virus: Methods and Protocols. ed. / Haitao Guo; Andrea Cuconati. Vol. 2837 2. ed. Netherlands: Springer, 2024. p. 137-148 (Methods in molecular biology (Clifton, N.J.)).

Research output: A Conference proceeding or a Chapter in Book › Chapter › peer-review

TY - CHAP

T1 - A Chromogenic In Situ Hybridization (CISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Liver Tissue

AU - Zhang, Xiaonan

AU - Lu, Wei

AU - Feng, Yanling

AU - Zhang, Zhanqing

AU - Yuan, Zhenghong

N1 - Publisher Copyright: © 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature. Funding Information: This work is supported by the Chinese National Natural Science Foundation (81873962, 32070152) and by Australian Centre for HIV and Hepatitis Virology Research (ACH2 ). Publisher Copyright: © The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024.

PY - 2024

Y1 - 2024

N2 - 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.

AB - 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.

KW - cccDNA

KW - CISH

KW - HBV

KW - Immunohistochemistry

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SN - 9781071640265

VL - 2837

T3 - Methods in molecular biology (Clifton, N.J.)

SP - 137

EP - 148

BT - Hepatitis B Virus

A2 - Guo, Haitao

A2 - Cuconati, Andrea

PB - Springer

CY - Netherlands

ER -

A Chromogenic In Situ Hybridization (CISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Liver Tissue

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