| 引用本文： |
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张占卿,陆伟,张小楠,等.肝组织HBV总DNA和cccDNA阴性参考值的探讨[J].同济大学学报（医学版）,2011,32(5):27-35. [点击复制]
- ZHANG Zhan-qing,LU Wei,ZHANG Xiao-nan,et al.The negative reference cut-offs of intrahepatic hepatitis B virus total DNA and covalently closed circular DNA[J].同济大学学报（医学版）,2011,32(5):27-35. [点击复制]
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| 摘要: |
| 目的 探讨肝组织乙型肝炎病毒（hepatitis B virus,HBV）总DNA（total DNA,tDNA）和共价闭合环状DNA（covalently closed circular DNA,cccDNA）的阴性参考值,为进一步研究实时定量PCR检测肝组织HBV tDNA和cccDNA的应用价值奠定基础。方法 分别有88例HBsAg阳性/抗-HBc阳性、20例HBsAg阴性/抗-HBc阳性和19例HBsAg阴性/抗-HBc阴性慢性肝炎患者入选本研究。肝组织β-globin DNA、HBV tDNA和cccDNA采用实时荧光定量PCR检测,单个肝细胞HBV tDNA和cccDNA含量（copies/cell）=HBV tDNA实测值（copies）/β-globin DNA实测值（copies）和HBV cccDNA实测值（copies）/β-globin DNA实测值（copies）。肝组织HBVtDNA和cccDNA含量的计算单位定义为log10copies/106cell。统计分析采用SPSS 13.0软件。结果 肝组织HBVtDNA和cccDNA鉴别HBsAg阳性/抗-HBc阳性与HBsAg阴性/抗-HBc阴性慢性肝炎的ROC曲线下面积分别为0.909和0.891（P=0.000和0.000）,最佳截断值分别为5.772和4.883 log10copies/66cells;鉴别HBsAg阳性/抗-HBc阳性与HBsAg阴性/抗-HBc阳性慢性肝炎的ROC曲线下面积分别为0.893和0.857（P=0.000和0.000.）,最佳截断值分别为5.559和4.630 log10copies/106cells。就HBsAg阳性/抗-HBc阳性慢性肝炎而言,当肝组织HBV tDNA和cccDNA≥5.699和cccDNA≥4.699 log10copies/106cells时,肝组织HBV tDNA与cccDNA含量之间均呈显著正相关（P=0.000和0.000）;当肝组织HBV tDNA＜5.699和cccDNA＜4.699 log10copies/106cells时,肝组织HBV tDNA与cccDNA含量之间均无显著相关性（P=0.065和0.880）。结论 肝组织HBV tDNA＜5.699和cccDNA＜4.699 log10copies/106cells可能是合适的阴性参考值。 |
| 关键词: 乙型肝炎病毒 共价闭合环状DNA 肝组织 实时定量PCR 正常参考值 |
| DOI：10.3969/j.issn1008-0392.2011.05.007 |
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| 基金项目:国家“八六三”高技术研究发展计划生物和医药技术领域重大/重点基金(2006AA02A411);国家科技重大专项(2008ZX10002-006) |
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| The negative reference cut-offs of intrahepatic hepatitis B virus total DNA and covalently closed circular DNA |
| ZHANG Zhan-qing1,LU Wei1,ZHANG Xiao-nan2,CAO Jie1 |
| (1.The Second Department of Hepatology,Shanghai Public Health Clinical Center of Fudan University,Shanghai 201508,China;2.Scientific Research Center,Shanghai Public Health Clinical Center of Fudan University,Shanghai 201508,China) |
| Abstract: |
| Objective To determine the negative reference cut-offs for intrahepatic hepatitis B virus （HBV） total DNA（tDNA） and covalently closed circular DNA（cccDNA）.Methods Eighty eight patients with HBsAg-positive and anti-HBc-positive,20 patients with HBsAg-negative and anti-HBc-positive and 19 patients with HBsAg-negative and anti-HBc-negative chronic hepatitis were enrolled in the present study. Theβ-globin DNA,HBV tDNA and cccDNA in liver tissue were determined by real-time PCR.The content of single hepatocyte HBV tDNA（copies/cell）=HBV tDNA measured value（copies）/B-globin DNA measured value（copies）,and the content of single hepatocyte HBV cccDNA（copies/cell）=HBV cccDNA measured value（copies）/β-globin DNA measured value（copies）.The calculating unit of the content of intrahepatic HBV tDNA and cccDNA was log10 copies/106 cells.SPSS 13.0 software was used for statistical analyses.Results The areas under the curves of ROC of intrahepatic HBV tDNA and cccDNA for distinguishing HBsAg-positive and anti-HBc-positive chronic hepatitis from HBsAg-negative and anti-HBc -negative chronic hepatitis were 0.909 and 0.891（P=0.000 and 0.000）,respectively;and the optimal cut-offs were 5.772 and 4.883 log10 copies/106 cells,respectively.The areas under the curves of ROC of intrahepatic HBV tDNA and cccDNA for distinguishing HBsAg-positive and anti-HBc-positive chronic hepatitis from HBsAg-negative and anti-HBc-positive chronic hepatitis were 0.893 and 0.857（P=0.000 and 0.000）,respectively;and the optimal cut-offs were 5.559 and 4.630 log10 copies/106 cells,respectively.In HBsAg-positive and anti-HBc-positive chronic hepatitis,the content of intrahepatic HBV tDNA was positively correlated significantly with the content of intrahepatic HBV cccDNA when intrahepatic HBV tDNA≥5.699 and HBV cccDNA≥4.699 log10 copies/106 cells（P=0.000 and 0.000）;and the content of intrahepatic HBV tDNA was not correlated significantly with the content of intrahepatic HBV cccDNA when intrahepatic HBV tDNA＜5.699 and HBV cccDNA＜4.699 log10 copies/106 cells（P=0.065 and 0.880）. Conclusion Intrahepatic HBV tDNA＜5.699 and HBV cccDNA<4.699 log10 copies/106 cells might be the rational negative reference cut-offs. |
| Key words: hepatitis B virus covalently closed circular DNA liver tissue Real-Time PCR normal reference |
