乙型肝炎病毒感染不同时期外周血单个核细胞微小RNA表达变化及其意义  

The significance and the change of miRNAs expression profile of peripheral blood mononuclear cell in different period of HBV infection

作　　者：张静[1] 史进方[1] 高纯[1] 蒋敏[1] 王玉林[1] 甘建和[2] 江敏华[2] 顾国浩[1] 

机构地区：[1]苏州大学附属第一医院检验科,215006 [2]苏州大学附属第一医院感染科

出　　处：《中华检验医学杂志》2012年第6期544-549,共6页Chinese Journal of Laboratory Medicine

基　　金：国家“十二五”科技重大专项子课题资助项目（2012ZX10002004-008）

摘　　要：目的探讨微小RNA（microRNAs，miRNAs）在乙型肝炎病毒（HBV）感染不同时期表达谱的变化及其意义。方法建立检测miRNA的RNADNA嵌合探针液态芯片（xMAP）法，对其特异性、重复性、准确性进行评估；2010年10月至2011年10月收集苏州大学附属第一医院住院和门诊急性乙型肝炎、慢性乙型肝炎、乙型肝炎后肝硬化及肝癌患者各40例，健康对照40名；采用xMAP液态芯片技术检测外周血单个核细胞（PBMC）中miR-191、-223、-222、-145、-21、-31、-126、-20a、-372的表达水平，以miR-103为内参，将（靶miRNA分子平均荧光强度一对应本底平均荧光强度）／（miR-103平均荧光强度-对应本底平均荧光强度）的比值作为有效数据分析各组miRNAs表达特征，采用SPSS17．0统计软件，组间比较用单因素方差分析，若差异有统计学意义，用LSD-t法行组间两两比较。结果xMAP液态芯片法检测miRNAs特异性为100％；重复性实验证实高值参比品的检测信号CV值均小于5％，低值参比品的检测信号CV值均小于10％；准确性实验证实9种miRNA的回收率为（1004-5）％；miR-222（F=1．32，P〉0．05）、-191（F=1．98，P〉0．05）、-145（F=0．78，P〉0．05）、-21（F=0．64，P〉0．05）、-31（F=0．83，P〉0．05）、-372（F=1．75，P〉0．05）组间表达差异无统计学意义；miR-223组间表达差异有统计学意义（F=14．56，P〈0．05），在急性乙型肝炎组表达最高（15．37±4．01），肝癌组表达最低（6．91±3．18）；组间miR-126表达差异有统计学意义（F=17．43，P〈0．05），在健康对照组表达最高（6．33±2．75），肝癌组表达最低（2．38±1．07）；组间miR-20a表达差异有统计学意义（F=19．48，P〈0．05），健康对照组表达最低（0．33±0．18），肝癌组表达最高（0．81±0．24）。结论xMAP液态芯片法检测miRNA特异性强、准确度高、重复性好，适合大通量�Objective To explore the significance and the change of miRNAs expression profile in different period of HBV infection. Methods Establish the detection method of microRNA（miRNA） by RNA DNA probe liquid chip （flexible multi-analyte profiling, xMap） and estimate the specificity, repeatability and accuracy of this method. From October 2010 to October 2011, collect HBV infected patients＇ peripheral blood of the First affiliated hospital of SooChow University, including acute hepatitis B, chronic hepatitis B, hepatits B liver cirrhosis, liver cancer patients and health control, each group contains 40 cases. The levels of miR-191 ,-223 ,-222 ,-145 ,-21 ,-31 ,-126 ,-20a,-372 of peripheral blood mononuclear cell were detected by xMap liquid chiptechnology. The level of miR-103 was taken as reference. The ratio of （the mean fluorescence intensity of target miRNAs-corresponding backgroud mean fluorescence intensity）/（ the mean fluorescence intensity of miR-103-corresponding backgroud mean fluorescence intensity） as a valid data and analysis the characteristics of miRNAs expression. The SPSS 17.0 was used as statistical software. The single factor analysis of variance was used as the method to analysis group comparison, and make multiple comparison by the LSD-t method if the result with a significant difference. Results The specificity of xMAP liquid chip method to detect miRNAs was 100% ;The repeated experiment proved that the CV value was less than 5% in the high value reference miRNA test, less than 10% in the low value reference miRNAs test; the accruracy experiment proved that the recovery rate was （100 ± 5 ）% in the nine miRNAs. There were no statistically differences with miR-222 （ F = 1.32, P 〉 0. 05 ）,-191 （ F = 1.98, P 〉 0. 05 ）,-145 （ F = 0. 78, P〉0.05）,-21（F=0.64,P〉0.05）,-31（F=0.83,P〉0.05）,-372（F=1.75,P〉0.05）in different groups ; There was statistically significant differences in miR-223 （ F = 14. 56, P 〈 0. 05 ） among different groups, with the hi

关 键 词：肝炎病毒 乙型 白细胞 单核 微RNAS 肝炎 乙型 芯片分析技术 

分 类 号：R51[医药卫生—内科学]