Langon T, Fillon S, Pichoud C, Hantz O, Trépo C, Kay A
INSERM U271, 151, Lyon, France.
Res Virol. 1998 May-Jun;149(3):171-85. doi: 10.1016/s0923-2516(98)80035-6.
Based on the analysis of HDV genomes from different areas of the world, three genotypes of HDV have been identified. Genotype I is the most prevalent and widespread. Genotype II is represented by two isolates from Japan and Taiwan. Genotype III has been found only in the Amazonian basin where it is associated with a history of severe disease, fulminant hepatitis with microvesicular steatosis (spongiocytosis). We report here the cloning and the analysis of the complete viral genome from woodchuck serum-derived HDV RNA after transmission from Central African Republic (RCA) patients with fulminant spongiocytic delta hepatitis. Two overlapping cDNA fragments, covering the entire HDV genome, were generated by RT-PCR and cloned. Three clones obtained from each fragment were fully sequenced. A complete consensus RCA HDV genome was reconstituted. The individual and the consensus nucleotide sequences were compared with those of 16 other fully sequenced isolates belonging to the three genotypes. Phylogenetic trees generated by the neighbour joining method firmly place our isolate in genotype I, and show that this RCA isolate differs significantly from the east African isolates previously analysed. Transfection experiments showed that the isolate is replication-competent, but less so than the control "wild-type" strain. Two novel mutations encountered in this work, one in the antigenomic ribozyme sequence and one affecting delta antigen, were studied.
基于对来自世界不同地区的丁型肝炎病毒（HDV）基因组的分析，已鉴定出HDV的三种基因型。基因型I最为普遍和广泛。基因型II由来自日本和台湾的两个分离株代表。基因型III仅在亚马逊河流域被发现，在那里它与严重疾病的病史相关，即伴有微泡脂肪变性（海绵样变）的暴发性肝炎。我们在此报告了从患有暴发性海绵样变丁型肝炎的中非共和国（RCA）患者传播后，从土拨鼠血清来源的HDV RNA中克隆并分析完整病毒基因组的情况。通过逆转录聚合酶链反应（RT-PCR）产生了覆盖整个HDV基因组的两个重叠互补DNA（cDNA）片段并进行了克隆。从每个片段获得的三个克隆进行了全序列测定。重建了一个完整的RCA HDV基因组共有序列。将个体和共有核苷酸序列与属于三种基因型的其他16个已完全测序的分离株的序列进行了比较。通过邻接法生成的系统发育树明确将我们的分离株归为基因型I，并表明该RCA分离株与先前分析的东非分离株有显著差异。转染实验表明该分离株具有复制能力，但比对照“野生型”菌株的复制能力弱。研究了在这项工作中遇到的两个新突变，一个在反基因组核酶序列中，另一个影响丁型抗原。
