乙型肝炎患者外周血单个核细胞中共价闭合环状DNA的检测及其临床意义  

Detection of the covalently closed circular DNA in peripheral blood mononuclear cells of hepatitis B patients and its clinical significance

作　　者：朱圣涛 谢琴秀[1] 张亚飞[1] 江晓平[1] 胡向阳[2] 柯晶[1] 李旭[1] 

机构地区：[1]安徽医科大学第一附属医院感染病科,合肥230032 [2]安徽医科大学第一附属医院病理科,合肥230032

出　　处：《中华传染病杂志》2014年第4期214-218,共5页Chinese Journal of Infectious Diseases

基　　金：国家自然科学基金资助项目（81273142）

摘　　要：目的 分析HBV感染者PBMC中HBV cccDNA与血清HBV DNA、HBsAg、HBeAg、肝组织学的相关性,并探讨其临床意义.方法 选取108例HBV感染者,密度梯度离心法提取PBMC,实时荧光定量PCR检测PBMC中HBV cccDNA和血清HBV DNA,化学发光定量检测血清中HBsAg和HBeAg,其中59例慢性HBV感染者行肝活组织检查.计数资料比较采用x2检验,计量资料比较采用相关性分析,计量资料不符合正态分布的采用非参数检验.结果 108例HBV感染者PBMC中HBV cccDNA阳性59例,占54.6％,其中15例肝功能衰竭患者HBV cccDNA阳性11例,明显高于8例急性乙型肝炎中2例阳性（x2 =4.960,P＜0.05）.根据患者血清HBV DNA载量分为＞5 lg、3～5 lg和＜3 lg拷贝/mL组,其PBMC中HBV cccDNA阳性率分别为76.1％（51/67）、5/18和13.0％（3/23）,HBV DNA高载量组与中、低载量组比较,差异有统计学意义（x2值分别为14.751、28.384,均P＜0.05）.PBMC中HBVcccDNA与血清HBV DNA载量和HBsAg呈正相关（r值分别为0.554、0.497,均P＜0.05）.PBMC中HBV cccDNA在肝组织学≥G2和（或）S2组检出率明显高于＜G2/S2组（x2=9.159,P＜0.05）.结论 HBV感染者PBMC中存在HBV cccDNA,且与血清HBV DNA和HBsAg呈正相关,也与肝组织学损伤相关.Objective To analyze the correlation between covalently closed circular DNA （cccDNA） in the peripheral blood mononuclear cells （PBMC） of hepatitis B virus （HBV）-infected patients and serum HBV DNA,hepatitis B surface antigen （HBsAg）,hepatitis B e antigen （HBeAg） and liver histology of hepatitis B patients,and to explore the clinical significance of HBV cccDNA detection in PBMC.Methods One hundred and eight patients with chronic HBV infection were involved in this study.PBMC were extracted using density gradient centrifugation.HBV cccDNA in PBMC and serum HBV DNA were detected by real-time fluorescence quantitative polymerase chain reaction.HBsAg and HBeAg were detected by chemiluminescence immunoassay.Liver biopsy was conducted in 59 out of the 108 patients.Chi-square test was used to compare the categorical variables.Correlation analysis was used to compare quantitative variables.Nonparametric test was used to compare the non-normal distribution parameters.Results In the overall population,HBV cccDNA in PBMC was positive in 59 patients （54.6%）.Eleven of the 15 patients with liver failure were found to be HBV cccDNA positive,which was significantly higher than that in the acute hepatitis B group （only 2 of the 8 patients were HBV cccDNA positive; x2 =4.960,P〈0.05）.One hundred and eight patients were categorized into three groups according to their serum HBV DNA levels,with group A：〉5 lg copy/mL,group B：3-5 lg copy/mL and group C：〈3 lg copy/mL.The proportions of HBV cccDNA positivity in PBMC in three groups were 76.1% （51/67）,5/18 and 13.0% （3/23）,respectively.Comparing with patients with lower HBV DNA （group B and C）,the proportion of HBV cccDNA positivity was higher in patients with higher HBV DNA （group A; x2=14.751,P〈0.05 and x2 =28.384,P〈0.05,resepectively）.The HBV cccDNA quantitation in PBMC was positively correlated with the serum HBV DNA level and HBsAg quantification （r=0.554,P〈0.05 and r=0.497,P〈0.05,respectively）.The proportion of HBV c

关 键 词：肝炎 乙型 慢性 肝炎病毒 乙型 离心法 梯密度 外周血单个核细胞 DNA 环状 

分 类 号：R512.62[医药卫生—内科学]