AIM: To investigate the frequency of mutations in pre-core (pre-C) and basic core promoter (BCP) regions of hepatitis B virus (HBV) from Shanxi Province, and the association between mutations and disease related index...AIM: To investigate the frequency of mutations in pre-core (pre-C) and basic core promoter (BCP) regions of hepatitis B virus (HBV) from Shanxi Province, and the association between mutations and disease related indexes.METHODS: One hundred chronic hepatitis B patients treated at Shanxi Province Hospital of Traditional Chinese Medicine were included in this study. PCR-reverse dot blot hybridization and mismatch amplification mutation assay (MAMA)-PCR were used to detect the mutations in the HBV pre-C and BCP regions. HBV DNA content and liver function were compared between patients with mutant HBV pre-C and BCP loci and those with wild-type loci. The consistency between PCR-reverse dot blot hybridization and MAMA-PCR for detecting mutations in the HBV pre-C and BCP regions was assessed.RESULTS: Of the 100 serum samples detected, 9.38% had single mutations in the pre-C region, 29.17% had single mutations in the BCP region, 41.67% had mutations in both BCP and pre-C regions, and 19.79% had wild-type loci. The rates of BCP and pre-C mutations were 65.7% and 34.3%, respectively, in hepatitis B e antigen (HBeAg) positive patients, and 84.6% and 96.2%, respectively, in HBeAg negative patients. The rate of pre-C mutations was significantly higher in HBeAg negative patients than in HBeAg positive patients (χ<sup>2</sup> = 26.62, P = 0.00), but there was no significant difference in the distribution of mutations in the BCP region between HBeAg positive and negative patients (χ<sup>2</sup> = 2.43, P = 0.12). The presence of mutations in the pre-C (Wilcoxon W = 1802.5, P = 0.00) and BCP regions (Wilcoxon W = 2906.5, P = 0.00) was more common in patients with low HBV DNA content. Both AST and GGT were significantly higher in patients with mutant pre-C and BCP loci than in those with wild-type loci (P < 0.05). PCR-reverse dot blot hybridization and MAMA-PCR for detection of mutations in the BCP and pre-C regions had good consistency, and the Kappa values obtained were 0.91 and 0.58, respectively.CONCLUSION: HBeAg negative patients tend to have HBV pre-C mutations. However, these mutations do not cause increased DNA copies, but associate with damage of liver function.展开更多
AIM： To explore mutations around the interferon sensitivity-determining region （ISDR） which are associated with the resistance of hepatitis C virus lb （HCV-lb） to interferon-α treatment. METHODS： Thirty-seven H...AIM： To explore mutations around the interferon sensitivity-determining region （ISDR） which are associated with the resistance of hepatitis C virus lb （HCV-lb） to interferon-α treatment. METHODS： Thirty-seven HCV-lb samples were obtained from Hong Kong patients who had completed the combined interferon-α/ribavirin treatment for more than one year with available response data. Nineteen of them were sustained virological responders, while 18 were non-responders. The amino acid sequences of the extended ISDR （eISDR） covering 64 amino acids upstream and 67 amino acids downstream from the previously reported ISDR were analyzed. RESULTS： One amino acid variation （I2268V, P = 0.023） was significantly correlated with treatment outcome in this pilot study with a limited number of patients, while two amino acid variations （R2260H, P = 0.05 and $2278T, P = 0.05） were weakly associated with treat- ment outcome. The extent of amino acid variations within the ISDR or eISDR was not correlated with treat- ment outcome as previously reported. CONCLUSION： Three amino acid mutations near but outside of ISDR may associate with interferon treatment resistance of HCV-lb patients in Hong Kong.展开更多
There are cases of hepatitis involving occult hepatitis B virus(HBV)infection in which,even though the HB surface antigen(HBsAg)is negative,HBV-DNA is detected by a polymerase chain reaction(PCR).We con-ducted a seque...There are cases of hepatitis involving occult hepatitis B virus(HBV)infection in which,even though the HB surface antigen(HBsAg)is negative,HBV-DNA is detected by a polymerase chain reaction(PCR).We con-ducted a sequence analysis of the entire HBV region in a case of non-B non-C chronic hepatitis in a 46-yearold female.A diagnosis of non-B non-C chronic hepatitis was made.Although HBV markers,such as HBs antibody(anti-HBs),anti-HBc,HBeAg and anti-HBe,were negative,HBV-DNA was positive.Nested PCR was performed to amplify the precore region of HBV-DNA and all remaining regions by long nested PCR.Sequence analysis of the two obtained bands was conducted by direct sequencing.Compared with the control strains,the ATG(Methionine)start codon in the X region had mut ated to GTG(Valine).It is assumed that a mutation at the start codon in the X region may be the reason why HBV markers are negative in some cases of hepatitis that involve occult HBV infection.展开更多
To determine the number of mutations in the NS5A region of the hepatitis C virus (HCV) and its relationship to the response to antiviral therapy in patients with chronic hepatitis C genotype 1 who are non-responders t...To determine the number of mutations in the NS5A region of the hepatitis C virus (HCV) and its relationship to the response to antiviral therapy in patients with chronic hepatitis C genotype 1 who are non-responders to two or more treatments. METHODSSequences within HCV NS5A [PKR binding domain (PKRBD) and the interferon-sensitivity-determining region (ISDR)] were analysed via direct sequencing in a selected cohort of 72 patients, with a total of 201 treatments [interferon-alpha (IFN-α), n = 49; IFN-α + ribavirin (RBV), n = 75; pegylated (peg) IFN-α + RBV, n = 47; first-generation direct-acting antivirals (DAAs), n = 13; and second-generation DAAs, n = 17]. Of these, 48/201 achieved a sustained virological response (SVR) and 153/201 achieved no virological response (NVR). RESULTSFor both regions, treatments resulting in SVR were associated with more baseline mutations than were treatments resulting in NVR (SVR vs NVR; PKRBD: 5.82 ± 3 vs 4.86 ± 2 mutations, P = 0.045; ISDR: 2.65 ± 2 vs 1.51 ± 1.7 mutations, P = 0.005). A decrease or no change in the number of mutations over time between treatments in the PKRBD or ISDR, as shown by sequencing, was associated with patients who usually failed to respond to treatment (PKRBD, P = 0.02; ISDR, P = 0.001). Moreover, patients showing a post-treatment baseline viral load > 600000 IU/mL and increased ISDR mutations with respect to the previous treatment were 9.21 times more likely to achieve SVR (P = 0.001). CONCLUSIONThe obtained results show that among patients who have shown no response to two or more antiviral treatments, the likelihood of achieving SVR increases with the genetic variability in the ISDR region (≥ 2 mutations or number of substitutions from the HCV-J and HCV-1 prototype), especially when the viral load is greater than 600000 IU/mL.展开更多
目的:研究扬州地区乙型肝炎病毒(hepatitis B virus,HBV)C基因区的基因特征,分析不同基因型间突变差异,为临床提供参考。方法:扩增HBV核酸C基因区并测序,Clustal W比对构建距离矩阵,MegAlign比对突变位点,分析不同基因型间核苷酸同源性...目的:研究扬州地区乙型肝炎病毒(hepatitis B virus,HBV)C基因区的基因特征,分析不同基因型间突变差异,为临床提供参考。方法:扩增HBV核酸C基因区并测序,Clustal W比对构建距离矩阵,MegAlign比对突变位点,分析不同基因型间核苷酸同源性和相似度;比较不同基因型间突变比例和突变频率间的差异,分析引起血清转换的突变。结果:测序成功31条序列,基因B型20例,C型11例,总突变率为77.4%,同源相似度分别为:96.1%~99.7%和98.1%~100.0%;基因B型有2例核苷酸G1896A和G1899A联合突变;氨基酸替换频率分别为:8.7/序列、3.3/序列;不同基因型突变比例和氨基酸替换频率的差异有统计学意义(χ^2=5.10,P=0.023,95%CI:1.14~49.26;χ^2=21.65,P<0.001,95%CI:1.816~4.650)。结论:扬州地区HBV C基因区同源相似度高,遗传距离小;HBV突变率高,存在产生血清学转换的核苷酸联合突变株,对病毒免疫学检验产生影响。展开更多
目的研究乙型肝炎病毒(hepatitis B virus,HBV)preC/C区基因突变与原发性肝癌患者预后的关系。方法收集81例乙型肝炎病毒相关性肝细胞肝癌(hepatitis B virus associated hepatocellular carcinoma,HBV-HCC)患者的癌组织并提取基因组DNA...目的研究乙型肝炎病毒(hepatitis B virus,HBV)preC/C区基因突变与原发性肝癌患者预后的关系。方法收集81例乙型肝炎病毒相关性肝细胞肝癌(hepatitis B virus associated hepatocellular carcinoma,HBV-HCC)患者的癌组织并提取基因组DNA,对HBVpreC/C区基因进行扩增和测序,并根据NCBI数据库鉴定出其突变位点,运用Kaplan-Meier和Cox回归等方法分析HBV-HCC患者的临床资料、突变位点与其术后生存期之间的关系。结果门脉瘤栓、肿瘤分期和肿瘤大小是与HBV-HCC患者术后生存相关的独立危险因素。1915、2134、2176、2221和2260突变位点被确定为预测HBV-HCC患者生存相关的独立危险因子,1979和2245突变位点与HBV-HCC患者生存具有临界统计学差异。结论门脉瘤栓、肿瘤分期和肿瘤大小以及HBVpreC/C区基因的以上7个突变位点被确定为与肝癌患者术后预后相关的独立危险因素。展开更多
目的探讨慢性乙型肝炎患者中拉米夫定诱导的 YMDD 变异和前 C 区1896位核苷酸以及 C区启动子1762和1764位核苷酸变异的检出率及其临床意义。方法采用基因芯片技术检测拉米夫定治疗6个月以上的122例慢性乙型肝炎患者血清中前 C 区1896位...目的探讨慢性乙型肝炎患者中拉米夫定诱导的 YMDD 变异和前 C 区1896位核苷酸以及 C区启动子1762和1764位核苷酸变异的检出率及其临床意义。方法采用基因芯片技术检测拉米夫定治疗6个月以上的122例慢性乙型肝炎患者血清中前 C 区1896位以及 C 区启动子1762和1764位核苷酸变异发生率。结果 122俐慢性乙型肝炎患者中检出40例 YMDD 变异阳性患者,检出率为32.8%。发生 YMDD变异后,HBV DNA 反跳,ALT 和 AST 增高,差异有统计学意义(P<0.01)。YMDD 变异阳性患者前 C 区1896位和 C 区启动子1762和1764位核苷酸变异检出率显著高于无 YMDD 变异的慢性乙型肝炎患者,差异有统计学意义(P<0.01)。YMDD 变异阳性伴前 C 区1896位以及 C 区启动子1762和1764位核苷酸变异患者与无前 C 区1896位以及 C 区启动于1762和1764位核苷酸变异的 YMDD 变异阳性患者比较,病情容易加重.易发展为重型肝炎或肝硬化,但差异无统计学意义(P>0.05)。结论拉米夫定诱导的 YMDD 变异患者容易发生前 C 区1896位/C 区启动子1762和1764位核菅酸变异,但与病情加重和预后无相关性。展开更多
基金Supported by Youth Foundation of Health and Family Planning Commission of Shanxi ProvinceNo.201301024
文摘AIM: To investigate the frequency of mutations in pre-core (pre-C) and basic core promoter (BCP) regions of hepatitis B virus (HBV) from Shanxi Province, and the association between mutations and disease related indexes.METHODS: One hundred chronic hepatitis B patients treated at Shanxi Province Hospital of Traditional Chinese Medicine were included in this study. PCR-reverse dot blot hybridization and mismatch amplification mutation assay (MAMA)-PCR were used to detect the mutations in the HBV pre-C and BCP regions. HBV DNA content and liver function were compared between patients with mutant HBV pre-C and BCP loci and those with wild-type loci. The consistency between PCR-reverse dot blot hybridization and MAMA-PCR for detecting mutations in the HBV pre-C and BCP regions was assessed.RESULTS: Of the 100 serum samples detected, 9.38% had single mutations in the pre-C region, 29.17% had single mutations in the BCP region, 41.67% had mutations in both BCP and pre-C regions, and 19.79% had wild-type loci. The rates of BCP and pre-C mutations were 65.7% and 34.3%, respectively, in hepatitis B e antigen (HBeAg) positive patients, and 84.6% and 96.2%, respectively, in HBeAg negative patients. The rate of pre-C mutations was significantly higher in HBeAg negative patients than in HBeAg positive patients (χ<sup>2</sup> = 26.62, P = 0.00), but there was no significant difference in the distribution of mutations in the BCP region between HBeAg positive and negative patients (χ<sup>2</sup> = 2.43, P = 0.12). The presence of mutations in the pre-C (Wilcoxon W = 1802.5, P = 0.00) and BCP regions (Wilcoxon W = 2906.5, P = 0.00) was more common in patients with low HBV DNA content. Both AST and GGT were significantly higher in patients with mutant pre-C and BCP loci than in those with wild-type loci (P < 0.05). PCR-reverse dot blot hybridization and MAMA-PCR for detection of mutations in the BCP and pre-C regions had good consistency, and the Kappa values obtained were 0.91 and 0.58, respectively.CONCLUSION: HBeAg negative patients tend to have HBV pre-C mutations. However, these mutations do not cause increased DNA copies, but associate with damage of liver function.
基金Supported by The National Key Technology Research and Development Program of China, No. 2009ZX10004-104
文摘AIM： To explore mutations around the interferon sensitivity-determining region （ISDR） which are associated with the resistance of hepatitis C virus lb （HCV-lb） to interferon-α treatment. METHODS： Thirty-seven HCV-lb samples were obtained from Hong Kong patients who had completed the combined interferon-α/ribavirin treatment for more than one year with available response data. Nineteen of them were sustained virological responders, while 18 were non-responders. The amino acid sequences of the extended ISDR （eISDR） covering 64 amino acids upstream and 67 amino acids downstream from the previously reported ISDR were analyzed. RESULTS： One amino acid variation （I2268V, P = 0.023） was significantly correlated with treatment outcome in this pilot study with a limited number of patients, while two amino acid variations （R2260H, P = 0.05 and $2278T, P = 0.05） were weakly associated with treat- ment outcome. The extent of amino acid variations within the ISDR or eISDR was not correlated with treat- ment outcome as previously reported. CONCLUSION： Three amino acid mutations near but outside of ISDR may associate with interferon treatment resistance of HCV-lb patients in Hong Kong.
文摘There are cases of hepatitis involving occult hepatitis B virus(HBV)infection in which,even though the HB surface antigen(HBsAg)is negative,HBV-DNA is detected by a polymerase chain reaction(PCR).We con-ducted a sequence analysis of the entire HBV region in a case of non-B non-C chronic hepatitis in a 46-yearold female.A diagnosis of non-B non-C chronic hepatitis was made.Although HBV markers,such as HBs antibody(anti-HBs),anti-HBc,HBeAg and anti-HBe,were negative,HBV-DNA was positive.Nested PCR was performed to amplify the precore region of HBV-DNA and all remaining regions by long nested PCR.Sequence analysis of the two obtained bands was conducted by direct sequencing.Compared with the control strains,the ATG(Methionine)start codon in the X region had mut ated to GTG(Valine).It is assumed that a mutation at the start codon in the X region may be the reason why HBV markers are negative in some cases of hepatitis that involve occult HBV infection.
基金Supported by“Consejería de Salud de la Junta de Andalucía”,No.PI0137/07“Instituto de Salud CarlosⅢ”,No.FISIntrasalud PI010/717
文摘To determine the number of mutations in the NS5A region of the hepatitis C virus (HCV) and its relationship to the response to antiviral therapy in patients with chronic hepatitis C genotype 1 who are non-responders to two or more treatments. METHODSSequences within HCV NS5A [PKR binding domain (PKRBD) and the interferon-sensitivity-determining region (ISDR)] were analysed via direct sequencing in a selected cohort of 72 patients, with a total of 201 treatments [interferon-alpha (IFN-α), n = 49; IFN-α + ribavirin (RBV), n = 75; pegylated (peg) IFN-α + RBV, n = 47; first-generation direct-acting antivirals (DAAs), n = 13; and second-generation DAAs, n = 17]. Of these, 48/201 achieved a sustained virological response (SVR) and 153/201 achieved no virological response (NVR). RESULTSFor both regions, treatments resulting in SVR were associated with more baseline mutations than were treatments resulting in NVR (SVR vs NVR; PKRBD: 5.82 ± 3 vs 4.86 ± 2 mutations, P = 0.045; ISDR: 2.65 ± 2 vs 1.51 ± 1.7 mutations, P = 0.005). A decrease or no change in the number of mutations over time between treatments in the PKRBD or ISDR, as shown by sequencing, was associated with patients who usually failed to respond to treatment (PKRBD, P = 0.02; ISDR, P = 0.001). Moreover, patients showing a post-treatment baseline viral load > 600000 IU/mL and increased ISDR mutations with respect to the previous treatment were 9.21 times more likely to achieve SVR (P = 0.001). CONCLUSIONThe obtained results show that among patients who have shown no response to two or more antiviral treatments, the likelihood of achieving SVR increases with the genetic variability in the ISDR region (≥ 2 mutations or number of substitutions from the HCV-J and HCV-1 prototype), especially when the viral load is greater than 600000 IU/mL.
文摘目的:研究扬州地区乙型肝炎病毒(hepatitis B virus,HBV)C基因区的基因特征,分析不同基因型间突变差异,为临床提供参考。方法:扩增HBV核酸C基因区并测序,Clustal W比对构建距离矩阵,MegAlign比对突变位点,分析不同基因型间核苷酸同源性和相似度;比较不同基因型间突变比例和突变频率间的差异,分析引起血清转换的突变。结果:测序成功31条序列,基因B型20例,C型11例,总突变率为77.4%,同源相似度分别为:96.1%~99.7%和98.1%~100.0%;基因B型有2例核苷酸G1896A和G1899A联合突变;氨基酸替换频率分别为:8.7/序列、3.3/序列;不同基因型突变比例和氨基酸替换频率的差异有统计学意义(χ^2=5.10,P=0.023,95%CI:1.14~49.26;χ^2=21.65,P<0.001,95%CI:1.816~4.650)。结论:扬州地区HBV C基因区同源相似度高,遗传距离小;HBV突变率高,存在产生血清学转换的核苷酸联合突变株,对病毒免疫学检验产生影响。
文摘目的研究乙型肝炎病毒(hepatitis B virus,HBV)preC/C区基因突变与原发性肝癌患者预后的关系。方法收集81例乙型肝炎病毒相关性肝细胞肝癌(hepatitis B virus associated hepatocellular carcinoma,HBV-HCC)患者的癌组织并提取基因组DNA,对HBVpreC/C区基因进行扩增和测序,并根据NCBI数据库鉴定出其突变位点,运用Kaplan-Meier和Cox回归等方法分析HBV-HCC患者的临床资料、突变位点与其术后生存期之间的关系。结果门脉瘤栓、肿瘤分期和肿瘤大小是与HBV-HCC患者术后生存相关的独立危险因素。1915、2134、2176、2221和2260突变位点被确定为预测HBV-HCC患者生存相关的独立危险因子,1979和2245突变位点与HBV-HCC患者生存具有临界统计学差异。结论门脉瘤栓、肿瘤分期和肿瘤大小以及HBVpreC/C区基因的以上7个突变位点被确定为与肝癌患者术后预后相关的独立危险因素。
文摘目的探讨慢性乙型肝炎患者中拉米夫定诱导的 YMDD 变异和前 C 区1896位核苷酸以及 C区启动子1762和1764位核苷酸变异的检出率及其临床意义。方法采用基因芯片技术检测拉米夫定治疗6个月以上的122例慢性乙型肝炎患者血清中前 C 区1896位以及 C 区启动子1762和1764位核苷酸变异发生率。结果 122俐慢性乙型肝炎患者中检出40例 YMDD 变异阳性患者,检出率为32.8%。发生 YMDD变异后,HBV DNA 反跳,ALT 和 AST 增高,差异有统计学意义(P<0.01)。YMDD 变异阳性患者前 C 区1896位和 C 区启动子1762和1764位核苷酸变异检出率显著高于无 YMDD 变异的慢性乙型肝炎患者,差异有统计学意义(P<0.01)。YMDD 变异阳性伴前 C 区1896位以及 C 区启动子1762和1764位核苷酸变异患者与无前 C 区1896位以及 C 区启动于1762和1764位核苷酸变异的 YMDD 变异阳性患者比较,病情容易加重.易发展为重型肝炎或肝硬化,但差异无统计学意义(P>0.05)。结论拉米夫定诱导的 YMDD 变异患者容易发生前 C 区1896位/C 区启动子1762和1764位核菅酸变异,但与病情加重和预后无相关性。
