摘要
目的对石蜡包埋肝组织核酸提取法进行优化,提高石蜡包埋肝组织乙型肝炎病毒总的去氧核糖核酸(HBV tDNA)、共价闭合环状去氧核糖核酸(cccDNA)定量检测的灵敏度,检测乙型肝炎(乙肝)相关肝癌患者的癌和癌旁组织中HBV tDNA、cccDNA。方法优化石蜡包埋肝组织核酸提取方法,采用荧光定量PCR法检测37例HBV感染相关肝细胞癌(HCC)患者石蜡包埋肝癌和癌旁组织中HBV tDNA、cccDNA。结果通过优化石蜡包埋肝组织核酸提取方法,核酸得量提高126.3%。HBV tDNA和cccDNA检出率在石蜡包埋肝癌组织中分别为100.0%(37/37)和97.3%(36/37),癌旁组织中均为100.0%(37/37)。HBV tDNA、cccDNA含量以及HBV cccDNA占tDNA的比例在肝癌组织与癌旁组织之间差异无统计学意义(P>0.05)。癌旁组织HBV tDNA与血清HBV DNA呈弱相关(r=0.380,P=0.020)。结论石蜡包埋肝组织核酸提取方法的优化可显著提高HBV相关的HCC患者癌和癌旁组织中HBV tDNA、cccDNA检出率。在癌旁组织中可100.0%检出HBV tDNA和cccDNA,建议对HCC切除术后的患者进行抗病毒治疗,以预防HBV感染相关HCC的复发。
Objective To optimize the method for nucleic acid extraction and to improve the sensitivity for quantitating HBV tDNA and cccDNA in formalin-fixed paraffin-embedded（FFPE）liver tissues.HBV tDNA and cccDNA in FFPE tumor and non-tumor liver tissues from HBV-related hepatocellular carcinoma（HCC）patients were detected.Methods The method for nucleic acid extraction was optimized and a real-time PCR assay was used for quantitating HBV tDNA and cccDNA in FFPE tumor and non-tumor liver tissues from 37 patients with HCC.Results The amount of extracted nucleic acid increased by 126.3% with the optimized method.The detection rates of HBV tDNA and cccDNA in FFPE tumor liver tissues were 100.0 %（37/37）and97.3%（36/37）,respectively;and the rates for both HBV tDNA and cccDNA in non-tumor liver tissues were100.0%（37/37）.There were no significant differences in intrahepatic HBV tDNA,cccDNA levels and the ratio of HBV cccDNA over tDNA between tumor and non-tumor tissues.The correlation analyses demonstrated that the serum HBV DNA level was weakly correlated with HBV tDNA level in non-tumor tissues（r=0.380,P=0.020）.Conclusions The detection rates of HBV tDNA and cccDNA in FFPE liver tissues were significantly increased when using the optimized method for the nucleic acid extraction.The HBV tDNA and cccDNA were detectable in the non-tumor liver tissues of all 37 patients.Antiviral therapy should be suggested for HCC patients with tumor resection to prevent tumor recurrence.
出处
《中国病毒病杂志》
CAS
2014年第3期203-209,共7页
Chinese Journal of Viral Diseases
基金
国家"十二五"艾滋病和病毒性肝炎等重大传染病防治科技重大专项(2013ZX10002004)
关键词
石蜡包埋肝组织
肝癌
乙型肝炎病毒
共价闭合环状DNA
定量检测
Formalin-fixed paraffin-embedded （FFPE） liver tissues
Hepatocellular carcinoma （HCC）
Hepa-titis B virus
Covalently closed circular DNA
Quantitative detection
