应用实时荧光定量RT.doc

应用实时荧光定量-PCR检测乙肝患者血浆中 miR-122的含量 黄琳淋，李光迪 通讯作者 通讯作者：李光迪，男，甘肃兰州人，兰州大学第二医院检验中心，主任检验师，Email:guangdi843133@163.com （兰州大学，第二临床医学院，甘肃 兰州 730030） 摘要： 目的 本实验主要检测miR-122在不同类型的乙型肝炎患者外周血浆中的含量，探询与天冬氨酸氨基转移酶、丙氨酸氨基转移酶的关系，明确血浆miR-122是否能作为新的乙肝肝损伤的标志物。方法 共收集80例血浆，其中乙肝患者60例，健康对照20例，采用实时荧光定量PCR检测miR-122和HBV-DNA，通过2-△△CT法计算各组血浆miR-122的相对量，标准曲线的绝对定量法检测各组血浆HBV-DNA含量；采用全自动生化分析仪检测天冬氨酸氨基转移酶（AST）及丙氨酸氨基转移酶(ALT)浓度。根据乙肝表面抗原情况分组，再依据HBV-DNA含量将乙肝表面抗原阳性组分层。结果 乙肝表面抗原阳性组的miR-122是乙肝表面抗原阴性组的25.16倍，AST及ALT在乙肝表面抗原阳性组的浓度【31.5(19-46.25)，30(23.25-44.75)】明显高于乙肝表面抗原阴性组【15(9.25-26)，18(16.25-23)】。将乙肝表面抗原阳性组分层，乙肝复制水平在7-8次方的miR-122含量水平明显高于复制水平在2-3次方及无复制层，差异有统计学意义（P=0.024，P=0.025）。miR-122与ALT间的相关系数为0.251，P=0.025，与AST间的相关系数为0.379，P=0.001，ALT与AST间的相关系数为0.821，P=0.00。结论 血浆miR-122浓度水平与乙肝病毒引起的肝损伤呈正相关。因而血浆miR-122有可能成为新的乙肝肝损伤标志物。 关键词 microRNA-122；天冬氨酸氨基转移酶；丙氨酸氨基转移酶；实时荧光定量RT-PCR；乙型肝炎 Detection of miR-122 expression in the plasma of patients with HBV- infected by real-time fluorescent quantitative-PCR Huang Linlin，Li Guangdi，Huang Kuo （The Second Clinical Medical College of Lanzhou Unicersity in Gansu province,Lanzhou 730030,China） Abstract：Objective We investigated whether miR-122 level in plasma differ in different groups of HBV-infected patients and how it changed.We also intend to clarify the relationship between miR-122 and Aspartate aminotransferase(AST),Alanine aminotransferase(ALT) and whether miR-122 in plasma would be a new marker for liver injure caused by hepatitis B virus.Method 80 cases were examinated including 60 cases with hepatitis B and 20 controls without hepatitis B.Real-time PCR was used for measuring the expression of miR-122 and HBV-DNA, 2 -△△CT method was used for calculated relative miR-122,standard curve method was calculated for HBV-DNA.Aspartate aminotransferase(AST) and Alanine aminotransferase(ALT) were detectived by full automatic biochemical analyzer.Depend on HBV surface antigen,cases was grouped into two groups,and HBV surface antigen-positive group was stratified by HBV-DNA.Results miR-12