AIM:To establish the more feasible and sensitive assessment approach to the detection of adefovir (ADV) resistance-associated hepatitis B virus (HBV) quasispecies.METHODS: Based on the characteristics of rtA181V/T and...AIM:To establish the more feasible and sensitive assessment approach to the detection of adefovir (ADV) resistance-associated hepatitis B virus (HBV) quasispecies.METHODS: Based on the characteristics of rtA181V/T and rtN236T mutations, a new approach based on real-time fluorescent quantitative polymerase chain reaction (RT-PCR) was established for the detection of ADV-resistant HBV quasispecies, total HBV DNA, rtA181 and rtN236 mutations in blood samples from 32 chronic hepatitis B (CHB) patients with unsatisfactory curative effect on ADV and compared with routine HBV DNA sequencing.RESULTS: Both the sensitivity and specificity of this new detection approach to ADV-resistant HBV quasispecies were 100%, which were much higher than those of direct HBV DNA sequencing. The approach was able to detect 0.1% of mutated strains in a total plasmid population. Among the 32 clinical patients, single rtA181 and rtN236T mutation and double rtA181T and rtN236T mutations were detected in 20 and 8, respectively, while ADV-resistant mutations in 6 (including, rtA181V/T mutation alone in 5 patients) and no associated mutations in 26.CONCLUSION: This new approach is more feasible and efficient to detect ADV-resistant mutants of HBV and ADV-resistant mutations before and during ADV treatment with a specificity of 100% and a sensitivity of 100%.展开更多
BACKGROUND Quantitative hepatitis B core-related antigen(qHBcrAg)has a better correlation with intrahepatic hepatitis B virus(HBV)covalently closed circular DNA(cccDNA)than HBV DNA or hepatitis B e antigen(HBeAg),but ...BACKGROUND Quantitative hepatitis B core-related antigen(qHBcrAg)has a better correlation with intrahepatic hepatitis B virus(HBV)covalently closed circular DNA(cccDNA)than HBV DNA or hepatitis B e antigen(HBeAg),but data are still lacking for its clinical application.AIM The aim was to investigate serum qHBcrAg levels in patients with chronic hepatitis B and assess the correlation of serum qHBcrAg with pregenomic RNA(pgRNA),cccDNA,and HBeAg seroconversion.METHODS This study was a secondary analysis of patients who underwent percutaneous liver biopsy between July 2014 and June 2019 in two multicenter randomized controlled clinical trials of peginterferon vs nucleos(t)ide analog(NUC)-based therapy(NCT03509688 and NCT03546530).Serum qHBcrAg,pgRNA,HBV DNA,hepatitis B core antigen,HBeAg,liver cccDNA,and HBV DNA were measured.The correlations of serum qHBcrAg with other biomarkers were analyzed.RESULTS A total of 139 patients were included.The mean qHBcrAg levels were 5.32±1.18 log10 U/mL at baseline and decreased during treatment(all P<0.0001).Serum qHBcrAg levels were positively correlated with pgRNA(r=0.597,P<0.0001)and cccDNA(r=0.527,P<0.0001)levels.The correlation of serum qHBcrAg level and intrahepatic HBV DNA levels at baseline was weak but significant(r=0.399,P<0.0001).HBcrAg predicted HBeAg seroconversion,with areas under the receiver operating characteristics curve of 0.788 at 24 wk and 0.825 at 48 wk.Log HBcrAg at wk 24 and 48 was independently associated with HBeAg seroconversion[odds ratio(OR)=2.402,95%confidence interval(CI):1.314-4.391,P=0.004;OR=3.587,95%CI:1.315-9.784,P=0.013].CONCLUSION Serum HBcrAg levels were correlated with HBV virological markers and could be used to predict HBeAg seroconversion.展开更多
AIMTo investigate potential predictors for treatment response to nucleos(t)ide analogues (NAs) in hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) patients. METHODSSeventy-six HBeAg-positive CHB patien...AIMTo investigate potential predictors for treatment response to nucleos(t)ide analogues (NAs) in hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) patients. METHODSSeventy-six HBeAg-positive CHB patients received 96-wk NAs optimized therapy (lamivudine and adefovir dipivoxil) were studied retrospectively. Serum hepatitis B surface antigen, HBeAg, hepatitis B core antibody, hepatitis B virus (HBV) DNA and alanine aminotransferase levels were quantitatively measured before and during the treatment at 12 and 24 wk. Stepwise logistic regression analyses were performed to identify predictors for treatment response, and areas under the receiver operating characteristic curves (AUROC) of the independent predictors were calculated. RESULTSForty-three CHB patients (56.6%) achieved virological response (VR: HBV DNA ≤ 300 copies/mL) and 15 patients (19.7%) developed HBeAg seroconversion (SC) after the 96-wk NAs treatment. The HBeAg level (OR = 0.45, P = 0.003) as well as its declined value (OR = 2.03, P = 0.024) at 24-wk independently predicted VR, with the AUROC of 0.788 and 0.736, respectively. The combination of HBeAg titer 1.6 lg PEIU/mL at 24-wk predicted VR with a sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) of 85%, 100%, 100% and 83%, respectively, and the AUROC increased to 0.923. The HBeAg level (OR = 0.37, P = 0.013) as well as its declined value (OR = 2.02, P = 0.012) at 24-wk also independently predicted HBeAg SC, with the AUROC of 0.828 and 0.814, respectively. The HBeAg titer 2.2 lg PEIU/mL at 24-wk predicted HBeAg SC with a sensitivity, specificity, PPV, NPV of 88%, 98%, 88% and 98%, respectively, and the AUROC reached 0.928. CONCLUSIONThe combination of HBeAg level and its declined value at 24-wk may be used as a reference parameter to optimize NAs therapy.展开更多
近年来,乙型肝炎e抗原(hepatitis B e Antigen,HBeAg)定量检测技术不断创新发展,由定性、半定量,发展至目前真正意义上的定量检测,并在临床上逐步推广应用。血清HBeAg定量水平与病毒复制、肝组织损伤以及谷氨酸-丙酮酸转氨酶(ALT)升高...近年来,乙型肝炎e抗原(hepatitis B e Antigen,HBeAg)定量检测技术不断创新发展,由定性、半定量,发展至目前真正意义上的定量检测,并在临床上逐步推广应用。血清HBeAg定量水平与病毒复制、肝组织损伤以及谷氨酸-丙酮酸转氨酶(ALT)升高等指标密切相关。基线HBeAg定量水平可预测聚乙二醇化干扰素(PEG-IFN)、核苷(酸)类似物(NAs)的疗效;治疗过程中实时监测HBeAg降低水平及幅度,可更加精准预测抗病毒药物的疗效,有助于指导临床医师针对不同患者制定个体化治疗方案。本文就目前HBeAg定量检测技术及其在临床应用方面的最新进展做一综述,旨在为临床医师判断患者病情变化,制定优化治疗方案等方面提供理论依据。展开更多
目的:探讨乙型肝炎病毒DNA(hepatitis B virus DNA,HBV-DNA)载量水平与血清学标志物(hepatitis B virus markers,HBVM)分组模式以及前S1抗原(Pre-S1 antigen,Pre-S1Ag)的关系。方法:收集2018年6月至2020年7月来院就诊的180例慢性乙肝患...目的:探讨乙型肝炎病毒DNA(hepatitis B virus DNA,HBV-DNA)载量水平与血清学标志物(hepatitis B virus markers,HBVM)分组模式以及前S1抗原(Pre-S1 antigen,Pre-S1Ag)的关系。方法:收集2018年6月至2020年7月来院就诊的180例慢性乙肝患者的血清样本,采用实时荧光定量聚合酶链反应(qRT-PCR)技术检测血清HBV-DNA载量水平;采用化学发光免疫分析法(CLIA)检测乙肝表面抗原(HBsAg)、e抗原(HBeAg)、表面抗体(抗HBs)、e抗体(抗HBe)、核心抗体(抗HBc),并归纳受检样本的HBVM分组模式;采用ELISA法检测血清Pre-S1Ag。分析HBV-DNA载量水平、HBVM分组模式和Pre-S1Ag水平的关系。结果:180例血清样本,HBsAg^(+)/抗HBe^(+)/抗HBc^(+)模式85例(47.22%),HBsAg^(+)/HBeAg^(+)/抗HBc^(+)模式70例(38.89%),其他模式25例(13.89%)。HBsAg^(+)/HBeAg^(+)/抗HBc^(+)模式组HBV-DNA、Pre-S1Ag阳性率均明显高于HBsAg^(+)/抗HBe^(+)/抗HBc^(+)模式组及其他模式组,差异有统计学意义(χ^(2)=56.955、46.809,P<0.05)。将HBV-DNA阳性作为判断HBV复制的金标准,HBeAg的灵敏度为87.23%,特异度为65.12%,阳性预测值为73.21%,阴性预测值为82.35%;Pre-S1Ag的灵敏度为90.35%,特异度为86.36%,阳性预测值为91.96%,阴性预测值为83.82%。依据HBV-DNA载量检测水平,分成<10^(3) copies/mL、10^(3)~10^(5) copies/mL、10^(5)~10^(7) copies/mL、>10^(7)copies/mL的4个亚组。随着HBV-DNA载量水平升高,Pre-S1Ag阳性率逐渐升高,分别为41.18%、64.00%、77.78%、94.29%,差异具有统计学意义(χ^(2)=31.250,P<0.05)。结论:HBV血清学标志物和Pre-S1Ag均可用于辅助诊断是否感染HBV以及HBV复制水平,但尚不可取代HBV-DNA定量检测,三者联合检测能为临床诊断、病毒复制提供更全面的依据。展开更多
基金Supported by The fund from Health Project of Jiangsu Province,No.H200711the AIDS,Hepatitis B and Other Infectious Diseases Prevention Program,No.2009ZX10004-712
文摘AIM:To establish the more feasible and sensitive assessment approach to the detection of adefovir (ADV) resistance-associated hepatitis B virus (HBV) quasispecies.METHODS: Based on the characteristics of rtA181V/T and rtN236T mutations, a new approach based on real-time fluorescent quantitative polymerase chain reaction (RT-PCR) was established for the detection of ADV-resistant HBV quasispecies, total HBV DNA, rtA181 and rtN236 mutations in blood samples from 32 chronic hepatitis B (CHB) patients with unsatisfactory curative effect on ADV and compared with routine HBV DNA sequencing.RESULTS: Both the sensitivity and specificity of this new detection approach to ADV-resistant HBV quasispecies were 100%, which were much higher than those of direct HBV DNA sequencing. The approach was able to detect 0.1% of mutated strains in a total plasmid population. Among the 32 clinical patients, single rtA181 and rtN236T mutation and double rtA181T and rtN236T mutations were detected in 20 and 8, respectively, while ADV-resistant mutations in 6 (including, rtA181V/T mutation alone in 5 patients) and no associated mutations in 26.CONCLUSION: This new approach is more feasible and efficient to detect ADV-resistant mutants of HBV and ADV-resistant mutations before and during ADV treatment with a specificity of 100% and a sensitivity of 100%.
基金by National Science and Technology Major Project,No.2017ZX10302201,No.2017ZX09304004 and No.2014ZX10002002National Program on Key Basic Research Project(973 Program),No.2015CB554304.
文摘BACKGROUND Quantitative hepatitis B core-related antigen(qHBcrAg)has a better correlation with intrahepatic hepatitis B virus(HBV)covalently closed circular DNA(cccDNA)than HBV DNA or hepatitis B e antigen(HBeAg),but data are still lacking for its clinical application.AIM The aim was to investigate serum qHBcrAg levels in patients with chronic hepatitis B and assess the correlation of serum qHBcrAg with pregenomic RNA(pgRNA),cccDNA,and HBeAg seroconversion.METHODS This study was a secondary analysis of patients who underwent percutaneous liver biopsy between July 2014 and June 2019 in two multicenter randomized controlled clinical trials of peginterferon vs nucleos(t)ide analog(NUC)-based therapy(NCT03509688 and NCT03546530).Serum qHBcrAg,pgRNA,HBV DNA,hepatitis B core antigen,HBeAg,liver cccDNA,and HBV DNA were measured.The correlations of serum qHBcrAg with other biomarkers were analyzed.RESULTS A total of 139 patients were included.The mean qHBcrAg levels were 5.32±1.18 log10 U/mL at baseline and decreased during treatment(all P<0.0001).Serum qHBcrAg levels were positively correlated with pgRNA(r=0.597,P<0.0001)and cccDNA(r=0.527,P<0.0001)levels.The correlation of serum qHBcrAg level and intrahepatic HBV DNA levels at baseline was weak but significant(r=0.399,P<0.0001).HBcrAg predicted HBeAg seroconversion,with areas under the receiver operating characteristics curve of 0.788 at 24 wk and 0.825 at 48 wk.Log HBcrAg at wk 24 and 48 was independently associated with HBeAg seroconversion[odds ratio(OR)=2.402,95%confidence interval(CI):1.314-4.391,P=0.004;OR=3.587,95%CI:1.315-9.784,P=0.013].CONCLUSION Serum HBcrAg levels were correlated with HBV virological markers and could be used to predict HBeAg seroconversion.
基金Supported by Major Science and Technology Special Project of China Twelfth Five-year Plan,Nos.2013ZX10002004 and 2012ZX10002003
文摘AIMTo investigate potential predictors for treatment response to nucleos(t)ide analogues (NAs) in hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) patients. METHODSSeventy-six HBeAg-positive CHB patients received 96-wk NAs optimized therapy (lamivudine and adefovir dipivoxil) were studied retrospectively. Serum hepatitis B surface antigen, HBeAg, hepatitis B core antibody, hepatitis B virus (HBV) DNA and alanine aminotransferase levels were quantitatively measured before and during the treatment at 12 and 24 wk. Stepwise logistic regression analyses were performed to identify predictors for treatment response, and areas under the receiver operating characteristic curves (AUROC) of the independent predictors were calculated. RESULTSForty-three CHB patients (56.6%) achieved virological response (VR: HBV DNA ≤ 300 copies/mL) and 15 patients (19.7%) developed HBeAg seroconversion (SC) after the 96-wk NAs treatment. The HBeAg level (OR = 0.45, P = 0.003) as well as its declined value (OR = 2.03, P = 0.024) at 24-wk independently predicted VR, with the AUROC of 0.788 and 0.736, respectively. The combination of HBeAg titer 1.6 lg PEIU/mL at 24-wk predicted VR with a sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) of 85%, 100%, 100% and 83%, respectively, and the AUROC increased to 0.923. The HBeAg level (OR = 0.37, P = 0.013) as well as its declined value (OR = 2.02, P = 0.012) at 24-wk also independently predicted HBeAg SC, with the AUROC of 0.828 and 0.814, respectively. The HBeAg titer 2.2 lg PEIU/mL at 24-wk predicted HBeAg SC with a sensitivity, specificity, PPV, NPV of 88%, 98%, 88% and 98%, respectively, and the AUROC reached 0.928. CONCLUSIONThe combination of HBeAg level and its declined value at 24-wk may be used as a reference parameter to optimize NAs therapy.
文摘近年来,乙型肝炎e抗原(hepatitis B e Antigen,HBeAg)定量检测技术不断创新发展,由定性、半定量,发展至目前真正意义上的定量检测,并在临床上逐步推广应用。血清HBeAg定量水平与病毒复制、肝组织损伤以及谷氨酸-丙酮酸转氨酶(ALT)升高等指标密切相关。基线HBeAg定量水平可预测聚乙二醇化干扰素(PEG-IFN)、核苷(酸)类似物(NAs)的疗效;治疗过程中实时监测HBeAg降低水平及幅度,可更加精准预测抗病毒药物的疗效,有助于指导临床医师针对不同患者制定个体化治疗方案。本文就目前HBeAg定量检测技术及其在临床应用方面的最新进展做一综述,旨在为临床医师判断患者病情变化,制定优化治疗方案等方面提供理论依据。
