Cancer Research Center of Lyon (CRCL), UMR Inserm 1052 CNRS 5286 Mixte, CLB, Université de Lyon 1 (UCBL1), Lyon, France.
Université Claude-Bernard Lyon I, Lyon, France.
Methods Mol Biol. 2024;2837:149-158. doi: 10.1007/978-1-0716-4027-2_13.
Chronic hepatitis B virus infection is a major health burden worldwide. Although efficient in maintaining the infection under control, current treatments are unable to fully eradicate the virus due to the persistence of its minichromosome, the so-called cccDNA. In the context of emerging antiviral and combinatorial therapies aiming at decreasing the cccDNA pool or at silencing its transcriptional activity, the detection and quantification of viral RNAs have gained increasing interest as a way to monitor the HBV reservoir. Here, we describe the protocol of the HBV full-length 5'RACE, a technique that allows to define the repertoire of the different HBV RNA species in vitro, intracellularly and extracellularly, and in vivo, in patients' serum in a qualitative or quantitative manner, depending on the choice of the post-analysis methodology.
慢性乙型肝炎病毒感染是全球范围内的一个主要健康负担。尽管目前的治疗方法能够有效地控制感染，但由于其微小染色体（cccDNA）的持续存在，无法完全清除病毒。在新兴的抗病毒和联合治疗方法的背景下，这些方法旨在减少 cccDNA 池或沉默其转录活性，病毒 RNA 的检测和定量作为监测 HBV 储存库的一种方法越来越受到关注。在这里，我们描述了 HBV 全长 5'RACE 的方案，该技术允许在体外、细胞内和细胞外以及体内以定性或定量的方式定义不同的 HBV RNA 种类的组合，具体取决于选择的分析后方法。
