为构建我国丁型肝炎病毒（HDV）全基因克隆，分别从3份HDV血清学阳性血清中提取病毒RNA，通过反转录-聚合酶链反应（RT-PCR）分段扩增，获得4个相互重叠的DNA片段，将PCR产物测序，利用DNAStar软件拼接，获得HDV全基因组序列；设计引物，用重叠PCR扩增全长HDV片段并连接至克隆载体，构建全基因克隆。结果成功克隆出3株1 675 bp的HDV全长基因组。经与GenBank标准序列比对，3株HDV均为基因Ⅰb型，核酸序列同源性达98%以上，与我国Ⅰ型X77627的同源性均达98%以上，与其他基因Ⅰ型的同源性均＞84%。与基因Ⅱ型和Ⅲ型的同源性分别低于77%和65%。本研究构建的3株具有我国代表性的HDV全长基因cDNA克隆，为进一步开展HDV分子生物学研究提供了基础。

The present paper aims to clone and analyze the sequence of full-length cDNA of hepatitis D virus (HDV) in China. HDV RNA was extracted from three HDV seropositive samples. cDNA was gotten by reverse transcription-polymerase chain reaction (RT-PCR). Four overlapped fragments were amplified by nested PCR, and the products were sequenced. The full-length sequences were joined by DNAStar. The full-length cDNA of HDV was amplified by overlapping PCR using designed primers and connected to the cloning T vector. Three strains of HDV were successfully amplified and cloned. The three HDV strains were all genotype I. The nucleic acid sequence homology of the three HDV strains was over 98%. The nucleic acid molecules of three strains had at least 84% sequence homology to genotype Ib. But they had over 98% sequence homology to genotype I Chinese strain X77627. They had no more than 77% and 65% sequence homology compared with genotype II and genotype III respectively. In conclusion, we cloned full-length cDNA of three strains of HDV, which could be helpful for the further study of HDV molecular biology.