实时荧光定量PCR检测乙型肝炎患者外周血单个核细胞CD_(58)基因的表达

Real-time Fluorescence Quantitative PCR in Detecting CD_(58) Gene Expression in PBMC in Patients with HBV Infection

目的:应用实时荧光定量PCR检测乙型肝炎患者外周血单个核细胞(PBMC)CD58基因的表达。方法:提取PBMC总RNA并逆转录为cDNA,实时荧光定量PCR法分别检测43例HBV感染者及11例正常对照的PBMCCD58基因的表达水平,同时检测血清ALT、AST水平。结果:乙型肝炎各组患者PBMCCD58mRNA表达水平较正常对照组明显增高,差异均有显著性。乙型肝炎患者PBMCCD58mRNA水平与血清ALT、AST呈显著正相关。结论:实时荧光定量PCR可以准确定量检测基因的表达;乙型肝炎患者PBMCCD58基因表达水平与疾病严重程度及肝细胞损伤严重程度有关。

Objective To quantitatively detect the expression level of CD_(58) in PBMC in the patients with HBV infection. Methods The total RNA isolated from PBMC was reversely transcribed into cDNA. The real-time fluorescence quantitative PCR method was used to analyze the expression level of CD_(58) gene in all patients and normal controls. The serum levels of ALT and AST were detected meanwhile. Results The real-time fluorescence quantitative PCR method was performed successfully to precisely detect the mRNA level. The expression level of CD_(58) gene in patients with HBV infection was significantly higher than those in control. The level of CD_(58) mRNA in PBMC from patients with HBV infection were positively correlated with serum levels of ALT and AST. Conclusion The real-time fluorescence quantitative PCR is the most precise method to quantitatively detect the mRNA level and can be used in gene expression changes. The expression level of CD_(58) gene is related with the degree of HBV infection and the degree of liver damage.