Persistent infection in patients with chronic hepatitis B proved

Subject to understand the HBV core gene promoter mutation and the degree of liver damage or HBeAg status of the relationship research purposes. Experiments with serum collected from the major hospital in Nanning in 59 patients with chronic hepatitis, 47 cases were male and 12 female patients aged 16 to 57 years. HBeAg-positive in 28 cases and negative in 31 cases. PCR reagents were purchased from the United States cloning technology labs, DNA purification reagents were purchased from the United States Promega Corporation, cycle sequencing kit purchased from the United States USB Corporation. DNA extracted by phenol / chloro-visit / iso-amyl alcohol (25/24/1) Conventional method extraction; DNA amplification: to do nested PCR, each round of amplification reaction conditions are the same: hot start 94 ℃ 5 minutes, then 94 ℃ 30 seconds, 55 ℃ 30 Miao, 72 ℃ 1.5 Fen Zhong. Continue to cycle after 30 cycles: 72 ℃ 10 Fenzhong, 22 ℃ 5 minutes; by direct sequencing of DNA sequencing.

Nested-PCR results showed that 59 cases of chronic hepatitis B patients 35 cases of HBV DNA positive, the positive rate was 59.3%. 16 patients with HBeAg-positive patients with HBV DNA positive, the positive rate of 57.1% (16/28), 19 patients with HBeAg-negative patients with HBV DNA positive, the positive rate of 61.3% (19/31). BCP sequencing results showed that without the usual sequence of samples, the most common type of mutation nt1762, 1764 pairs of sudden occurrence (A → T, G → A), accounting for 57.1% (20/35); followed by mutation types nt1799 sites mutation, from C → G, 54.4% (19/35), for non-sense mutation; nt1752 mutation, from the A → G, so that the password by isoleucine into valine, accounting for 37.1% (13 / 35); nt1753 T → C, accounting for 20.0% (7 / 35).

Core antigen and e antigen, some of the same sequence, this part of the sequence can be the host cytotoxic T lymphocytes recognize the core peptide of the changes in the immune attack may be caused by chronic liver disease. The results showed that samples of each district are base pairs in the Enh2/CP mutation, the most common mutation nt1762, 1764 pairs of sudden occurrence (A → T, G → A), meeting the common mutations are nt1799 mutation, from C → G, for the non-sense mutation; nt1752 mutation from A → G, so that the password by isoleucine into valine, nt1753 T → C. Core gene promoter to make hepatitis B virus mutations (HBV) can be persistent infection in the body, these mutations in patients with chronic active hepatitis related to.