Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology · 2000
[Cloning and sequencing of partial genes of hepatitis C virus genome in patients with acute hepatitis C].
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Jiang D, Yao D, Dong Z, Qiu L, Lu J
Аннотация
OBJECTIVE: To explore the etiological role of HCV in patients with acute hepatitis.
METHODS: The prevalence of HCV infection in 89 patients with acute hepatitis was investigated by analysis of HCV RNA and HCV second generation antibody. HCV RNAs extracted from the sera of 5 patients with NANB acute hepatitis, which were positive for HCV RNA, were converted to cDNA by reverse transcription with random primer and genotyping by PCR with type-specific primers. The partial genes of HCV genome were amplified. The PCR products were expressed in E. coli with p-GEM-T vector, and their nucleotide sequences were determined by dideoxynucleotide chain-termination method.
RESULTS: The incidence of hepatitis virus infection was 47. 2% in HAV, 28.1% in HBV and 15.7% in HCV, respectively. The incidence of HAV and HBV coinfection was 14.6% and the rate of non-A, non-B and non-C hepatitis was 9% in all patients. The genotype of HCV-RNA positive patients was 85.8% in HCV-II, 7.1% in HCV-III and 7. 1% in combining HCV-II/III, respectively. The partial sequence of HCV genome in 5 patients with non-A and non-B acute hepatitis was amplified and the fragment was 424 bp in accordance with original design. The homology of the sequences was 98.1%-99.5% in nucleotide acid and 97.6%-99.2% in amino acid among five isolates. The average homology was 91.9% or 94.3%-95.6% for nucleotide sequences between HCV-I or HCV-II and the 5 isolates, and 92.3%-95.8% for amino acid sequences between the 5 isolates and HCV-I or HCV-II, respectively.
CONCLUSION: HCV infection is one of the main hepatitis viruses in patients with acute hepatitis, in which the HCV-II genotype is dominant and should be paid attention to it.
METHODS: The prevalence of HCV infection in 89 patients with acute hepatitis was investigated by analysis of HCV RNA and HCV second generation antibody. HCV RNAs extracted from the sera of 5 patients with NANB acute hepatitis, which were positive for HCV RNA, were converted to cDNA by reverse transcription with random primer and genotyping by PCR with type-specific primers. The partial genes of HCV genome were amplified. The PCR products were expressed in E. coli with p-GEM-T vector, and their nucleotide sequences were determined by dideoxynucleotide chain-termination method.
RESULTS: The incidence of hepatitis virus infection was 47. 2% in HAV, 28.1% in HBV and 15.7% in HCV, respectively. The incidence of HAV and HBV coinfection was 14.6% and the rate of non-A, non-B and non-C hepatitis was 9% in all patients. The genotype of HCV-RNA positive patients was 85.8% in HCV-II, 7.1% in HCV-III and 7. 1% in combining HCV-II/III, respectively. The partial sequence of HCV genome in 5 patients with non-A and non-B acute hepatitis was amplified and the fragment was 424 bp in accordance with original design. The homology of the sequences was 98.1%-99.5% in nucleotide acid and 97.6%-99.2% in amino acid among five isolates. The average homology was 91.9% or 94.3%-95.6% for nucleotide sequences between HCV-I or HCV-II and the 5 isolates, and 92.3%-95.8% for amino acid sequences between the 5 isolates and HCV-I or HCV-II, respectively.
CONCLUSION: HCV infection is one of the main hepatitis viruses in patients with acute hepatitis, in which the HCV-II genotype is dominant and should be paid attention to it.