Chao Y C, Lee C M, Tang H S, Govindarajan S, Lai M M
Howard Hughes Medical Institute, University of Southern California, School of Medicine, Los Angeles 90033.
Hepatology. 1991 Feb;13(2):345-52.
The genomic RNA of an Asian isolate of hepatitis delta virus was cloned from a Chinese patient from Taiwan, using the polymerase chain reaction to amplify cDNA for cloning and sequencing. The sequence of this hepatitis delta virus isolate shares an 86% to 88% similarity with the three published hepatitis delta virus RNA sequences, suggesting heterogeneity of hepatitis delta viruses from different geographical areas. Four highly conserved, long stretches of sequence were found. These four regions corresponded to the sequences required for the autocatalytic cleavage activities of the genomic and antigenomic RNAs and the middle and the carboxyl terminal parts of the open reading frame for the delta antigen on the antigenomic strand. The conservation of nucleotide sequence in these four regions was further confirmed by sequencing additional hepatitis delta virus RNAs obtained from three patients with chronic delta hepatitis who lived in Los Angeles. These findings suggest that the conserved sequences are critical for viral replication. These conserved regions offer ideal sites for primer selection to carry out polymerase chain reactions to detect hepatitis delta virus RNA in patients with hepatitis delta virus infection.
从一名来自中国台湾的患者身上克隆出丁型肝炎病毒亚洲分离株的基因组RNA,采用聚合酶链反应扩增cDNA用于克隆和测序。该丁型肝炎病毒分离株的序列与已发表的三条丁型肝炎病毒RNA序列有86%至88%的相似性,提示来自不同地理区域的丁型肝炎病毒具有异质性。发现了四段高度保守的长序列。这四个区域对应于基因组和反基因组RNA的自催化切割活性以及反基因组链上丁型抗原开放阅读框的中部和羧基末端部分所需的序列。通过对另外三名居住在洛杉矶的慢性丁型肝炎患者的丁型肝炎病毒RNA进行测序,进一步证实了这四个区域核苷酸序列的保守性。这些发现提示保守序列对病毒复制至关重要。这些保守区域为引物选择提供了理想位点,可用于开展聚合酶链反应以检测丁型肝炎病毒感染患者的丁型肝炎病毒RNA。