乙型肝炎病毒前基因组RNA检测在乙型肝炎治疗中的临床研究

Clinical application of hepatitis B virus pregenomic RNA in diagnosis and treatment of patients with hepatitis B

目的探讨乙型肝炎病毒前基因组RNA(HBV-pgRNA)的检测在乙型肝炎治疗中的临床价值.方法选取怀化市第一人民医院2017年5月-2018年10月收治的乙型肝炎患者80例,采用定量聚合酶链式反应(qPCR)检测不同感染阶段及治疗期间患者血清HBV pgRNA、HBV DNA水平,用化学发光法检测血清乙肝表面抗原(HBsAg)滴度,肝活检组织中提取DNA后检测共价闭合环状DNA(cccDNA),分析HBV pgRNA与HBV DNA、HBsAg、cccDNA的相关性及预测患者治疗的价值.结果血清HBV pgRNA、HBV DNA在乙肝e抗原(HBeAg)阳性慢性HBV感染期(Ⅰ+Ⅱ)最高,HBeAg阴性慢性HBV感染期(Ⅲ+Ⅳ)最低,血清HBsAg水平和肝内cccDNA水平在四个阶段保持相对稳定.对总体患者、Ⅰ期和II期患者,HBV pgRNA与HBV DNA、HBsAg、cccDNA均成正相关(P<0.05);HBV pgRNA、HBV DNA、HBsAg、cccDNA水平治疗后均下降,除HB-sAg治疗后12个月与6个月无统计学差异外,其他各指标治疗后与前一治疗阶段差异均有统计学意义(P<0.05);HBV cccDNA水平(AUC=0.773)是HBsAg血清转换的最佳预测指标,其次是HBV pgRNA水平(AUC=0.743).HBV pgRNA水平对HBsAg血清转换预测敏感度89.11%,特异度81.23%,准确度85.72%.结论HBV pgRNA的动态变化可以作为一种新的标志物来预测HBeAg阳性患者治疗.

OBJECTIVE To explore the clinical value of hepatitis B virus pregenomic RNA(HBV-pgRNA)in treatment of patients with hepatitis B. METHODS A total of 80 patients with hepatitis B who were treated in Huaihua First People’s Hospital from May 2017 to Oct. 2018 were enrolled in the study,the levels of serum HBV pgRNA and HBV DNA of the patients were detected by using quantitative polymerase-chain-reaction(qPCR) at different stages of infection and during the treatment, the serum HBsAg titers were measured by chemiluminescence method at different stages of infection and during treatment, DNA was extracted from liver biopsy tissue so as to detect the covalently closed circular DNA(cccDNA), and the correlation between HBV pgRNA and HBV DNA, HBsAg and cccDNA as well as its value in prediction of treatment was observed. RESULTS The levels of serum HBV pgRNA and HBV DN were the highest among the hepatitis B eantigen(HBeAg)-positive patients during stage(Ⅰ+Ⅱ) of chronic HBV infection and were the lowest among the HBeAg-negative patients during stage(Ⅲ+Ⅳ) of chronic HBV infection, while the levels of serum HBsAg and intrahepatic cccDNA remained stable during the four states. As for the overall patients, patients with infection of stage I and patients with infection of stage II, the HBV pgRNA was positively correlated with HBV DNA, HBsAg and cccDNA(P<0.05). The levels of HBV pgRNA, HBV DNA, HBsAg and cccDNA were declined after the treatment, there was no significant difference in the HBsAg after the treatment for 12 and 6 months, while, there were significant differences in other indexes before and after the treatment for different time periods(P<0.05). The HBV cccDNA level(AUC=0.773) was the optimal predictor for seroconversion of HBsAg, followed by the HBV pgRNA level(AUC=0.743). The sensitivity of the HBV pgRNA was 89.11% in prediction of seroconversion of HBsAg, the specificity 81.23%, the accuracy 85.72%. CONCLUSION The dynamic change of HBV pgRNA can be used as an novel marker for prediction of the treatment of the HBeAg-positive patients.