title: [Complete HBV DNA clone and sequence from serum samples of severe hepatitis B patients].
abstract: OBJECTIVE
  To study the association between hepatitis B virus (HBV) mutants and the pathogenesis of severe hepatitis B by full-length HBV genome.
  
  
  METHODS
  Serum samples from 10 severe hepatitis B patients were collected in our hospital. Serum HBV DNAs were extracted using DNA mini Kit, and amplified by LA Taq DNA polymerase to yield full-length HBV DNA. PCR products were isolated and cloned into vector pUCm-T, then transfected into DH-5 alpha cells. Positive clones were selected and checked by digestion, and full-length HBV DNAs were sequenced.
  
  
  RESULTS
  4 cases were cloned into vector pUCm-T successfully and completed the full-length sequencing. Among them, 3 cases had a G to A mutation at nucleotide 1896 in pre-C region and 1 had a double mutation of T1762-A1764 in the core promoter region. Some amino acid changes occurred within the known CTL, B or T cell epitopes of the PrS2 and C regions.
  
  
  CONCLUSIONS
  This method could serve to study the relationship between HBV genome and the pathogenesis of severe hepatitis B.
authors.affiliation: Department of Infectious Diseases, First Hospital of Zhejiang University, Hangzhou 310003, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.forename: Lan-juan
authors.initials: LJ
authors.lastname: Li
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.forename: Xiang
authors.initials: X
authors.lastname: Qian
authors.forename: Dong-qing
authors.initials: DQ
authors.lastname: Cheng
mesh_terms: D000328:Adult; D001402:B-Lymphocytes / Q000378:metabolism; D003001:Cloning, Molecular; D000939:Epitopes / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics; D006509:Hepatitis B / Q000209:etiology* / Q000821:virology; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D011498:Protein Precursors / Q000235:genetics*; D017421:Sequence Analysis; D013602:T-Lymphocytes, Cytotoxic / Q000378:metabolism
chemical_list: D000939:Epitopes; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; C050807:presurface protein 2, hepatitis B surface antigen