Müller H M, Pfaff E, Goeser T, Theilmann L
Department of Internal Medicine, University of Heidelberg, Federal Republic of Germany.
J Med Virol. 1993 Aug;40(4):291-306. doi: 10.1002/jmv.1890400407.
Heterogeneity of hepatitis C viral (HCV) genomes of several isolates from different countries has been reported, but there is little information on HCV isolates for the Federal Republic of Germany. Therefore, the nucleotide (nt) and deduced amino acid (aa) sequences of interesting parts of the viral genome derived from different human isolates in Germany were compared with each other and with the nt and predicted aa sequences of recently published isolates. HCV sequences were obtained by reverse transcription of viral RNA extracted from serum followed by polymerase chain reaction (PCR) amplification. Within the 5' nontranslated region we found only 3 single nucleotide exchanges among 2 of our isolates, and in comparison to sequences of Japanese isolates 2 to 3 exchanges, and to U.S. isolates 1 to 5 exchanges (homologies 98% to > 99%). Determination of a 249-bp core sequence from two German isolates exhibited 3% sequence divergence. The sequence of the core region (nt 342-911) showed a homology of about 88-91% on nt level and 96-97% on aa level as compared to U.S. isolates and other German isolates, and a homology of 95-96% (nt) and 96-98% (aa), respectively, to Japanese isolates. Less homologies were noticed for the E1 and E2/NS1 genes, especially in the N-terminal E2/NS1 hypervariable domain. Our isolates HD1 and HD2 showed nt sequence homologies of about 72-81% and aa homologies of 76-88% to U.S., German, and French isolates, and 89-91% (nt) and 88-96% (aa), respectively, to Japanese isolates. These results indicate that various German isolates are more closely related to Japanese isolates and differ from other European isolates as reported so far. Because of a nucleotide sequence heterogeneity of up to 10% among the tested isolates, we conclude that more than one closely related but distinct viral genotype of HCV exists in Germany. Furthermore, heterogeneous sequences of HCV can be detected in a single patient suggesting multiple infection with different genomic variants or, alternatively, a genetic drift forced by mutational events as a consequence of host immune selection.
已有报道称来自不同国家的几种丙型肝炎病毒（HCV）分离株的基因组存在异质性，但关于德意志联邦共和国的HCV分离株的信息却很少。因此，将德国不同人类分离株的病毒基因组中有趣部分的核苷酸（nt）序列和推导的氨基酸（aa）序列相互进行了比较，并与最近公布的分离株的nt序列和预测的aa序列进行了比较。通过对从血清中提取的病毒RNA进行逆转录，然后进行聚合酶链反应（PCR）扩增来获得HCV序列。在5'非翻译区，我们在2株分离株中仅发现3个单核苷酸交换，与日本分离株的序列相比有2至3个交换，与美国分离株相比有1至5个交换（同源性为98%至>99%）。对两株德国分离株的249bp核心序列进行测定，显示出3%的序列差异。与美国分离株和其他德国分离株相比，核心区域（nt 342 - 911）的序列在nt水平上的同源性约为88 - 91%，在aa水平上为96 - 97%，与日本分离株的同源性分别为95 - 96%（nt）和96 - 98%（aa）。对于E1和E2/NS1基因，同源性较低，尤其是在E2/NS1 N端高变区。我们的分离株HD1和HD2与美国、德国和法国分离株的nt序列同源性约为72 - 81%，aa同源性为76 - 88%，与日本分离株的同源性分别为89 - 91%（nt）和88 - 96%（aa）。这些结果表明，各种德国分离株与日本分离株的关系更为密切，与迄今为止报道的其他欧洲分离株不同。由于在测试的分离株中核苷酸序列异质性高达10%，我们得出结论，德国存在不止一种密切相关但不同的HCV病毒基因型。此外，在单个患者中可以检测到HCV的异质序列，这表明存在不同基因组变体的多重感染，或者是由于宿主免疫选择导致的突变事件引起的基因漂移。
