Abstract：
The hepatitis delta virus (HDV) is a small RNA virus (1700 base pairs), which uses the surface proteins of the hepatitis B virus (HBV) as an envelope. Accurate and reliable quantitative detection of HDV RNA is central for scientific and translational clinical research or diagnostic purposes. However, HDV poses challenges for nucleic acid amplification techniques: (1) the circular genome displays high intramolecular base pairing; (2) high content of cytosine and guanine; and (3) enormous genomic diversity among the eight known HDV genotypes (GTs). Here, we provide step-by-step instructions for (A) a manual workflow to perform a quantitative HDV reverse transcription (RT)-PCR from serum and liver tissue and (B) a quantitative HDV RT-PCR assay with whole process control to be used for serum or plasma samples run on a fully automated system. Both assays target the conserved ribozyme region and demonstrate inclusivity for all eight HDV GTs. The choice of assay depends on the experimental needs and equipment availability. While the former is ideal for scientific research laboratories, the latter provides a useful tool in the field of translational research or diagnostics.
摘要：
丁型肝炎病毒（HDV）是一种小RNA病毒（1700个碱基对），它使用乙型肝炎病毒（HBV）的表面蛋白作为包膜。准确可靠的HDVRNA定量检测对于科学和转化临床研究或诊断目的至关重要。然而,HDV对核酸扩增技术提出了挑战:(1)环状基因组显示高度的分子内碱基配对;(2)高含量的胞嘧啶和鸟嘌呤;以及(3)在八种已知的HDV基因型(GT)中的巨大基因组多样性。这里,我们提供了以下分步说明:(A)从血清和肝组织进行定量HDV逆转录(RT)-PCR的手动工作流程;(B)在全自动系统上运行的具有全过程控制的定量HDVRT-PCR测定,用于血清或血浆样品.两种测定都靶向保守的核酶区域,并且证明了所有八个HDVGT的包容性。测定的选择取决于实验需要和设备可用性。虽然前者是科学研究实验室的理想选择，后者在转化研究或诊断领域提供了有用的工具。
