Real-time quantitative PCR analysis of human renal biopsies

Human biopsies were collected from kidney transplant recipients in 4 different european centers in the context of BIOMARGIN study. We used TaqMan™ Advanced miRNA Human A and B Cards to quantitate 754 miRNAs expression according to patient outcome ; ie patient presenting Microvascular Inflammation (MVI) or No MVI . qPCR MiRNA expression profiling. Needle core biopsies from 88 kidney transplant recipients were collected and immediatly stored in Allportect Tissue Reagent at -20°C before RNA extraction. Equal amount of total RNA (150 ng) from each patient was used to gene expression analysis.本研究依托BIOMARGIN研究项目，从欧洲4家不同医学中心的肾移植受者处采集人体活检样本。我们采用TaqMan™ Advanced miRNA人类A、B检测卡，针对患者结局（即出现微血管炎症（Microvascular Inflammation，MVI）或无MVI）对754种微小RNA（miRNA）的表达水平进行定量，并通过实时荧光定量PCR（qPCR）开展miRNA表达谱分析。本研究共收集88名肾移植受者的穿刺芯活检样本，在进行RNA提取前，即刻将样本置于-20℃的Allportect组织保存试剂中储存。从每位患者样本中提取等量的总RNA（150 ng），用于基因表达分析。