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Detection of Low Copy Number Integrated Viral DNA Formed by In Vitro Hepatitis B Infection

Detection of Low Copy Number Integrated Viral DNA Formed by In Vitro Hepatitis B Infection

Published on: November 7, 2018

一种多用途的方法来分析乙型肝炎病毒的DNA整合.

Kento Fukano^1,2, Kousho Wakae¹, Naganori Nao^3,4,5

¹Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.

Hepatology communications

|December 5, 2023

概括

一种名为RAISING的新方法有效地检测B型肝炎病毒 (HBV) 在宿主基因组中的DNA整合部位. 这种具有成本效益的技术有助于了解HBV相关的肝癌和诊断.

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科学领域:

分子生物学分子生物学
基因组学就是基因组学.
肝病学肝病学是一种肝病学.

背景情况:

肝细胞癌 (HCC) 通常与乙型肝炎病毒 (HBV) 的DNA整合到宿主基因组有关,这是肝癌发生的一个关键因素.
目前用于检测HBV融合部位的现有方法在灵敏度,成本和吞吐量方面存在局限性.
需要灵敏,全面和具有成本效益的方法来检测病毒集成部位.

研究的目的:

调查RAISING (快速扩大集成部位而无基因组DNA污染干扰) 检测HBV DNA集成的效率.
建立RAISING作为一种简单,廉价和高度敏感的方法来识别病毒集成.

主要方法:

在RAISING中,使用特定于病毒的原始剂来放大HBV集成的片段,覆盖整个HBV基因组.
使用Illumina对产品进行了测序,在HCC细胞系中确定了HBV-人类结序列.
在各种模型和临床标本中,使用结点特定的嵌套PCR和滴滴数字PCR进行了验证.

主要成果:

通过RAISING测序,成功地确定了HBV-人类结序列及其在HCC细胞系中的频率.
该方法在集中结合序列方面表现出优异的性能,与混合捕获序列相比.
在各种细胞类型,动物模型和临床样本中,RAISING测序准确地确定了de novo整合点.

结论:

RAISING提供了一种具有成本效益和灵敏的方法来分析HBV-人类结合序列,需要更少的测序数据.
预计这种方法将推动对基本HBV整合机制的研究.
而RAISING即将成为HBV相关疾病临床诊断的宝贵工具.