肺结核病患者外周血单个核细胞颗粒溶素基因的表达及临床意义  

Expression and clinical significance of granulysin in peripheral blood mononuclear cells of patients with pulmonary tuberculosis

作　　者：缪家文 张海燕 王捷婷 王雪梅 苏琴 潘洪秋 李芳秋[2] 

机构地区：[1]江苏省镇江市第三人民医院中心实验室,镇江市212005 [2]南京军区南京总医院解放军临床检验医学研究所

出　　处：《中华实验和临床感染病杂志（电子版）》2012年第5期22-25,共4页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：国家自然科学基金资助项目(No.30670599);镇江市科技支撑-社会发展项目(No.SH2011062)

摘　　要：目的建立检测人颗粒溶素(GNLY)mRNA含量的实时荧光定量RT-PCR的方法,并测定肺结核病患者外周血单个核细胞(PBMC)中GNLY的基因表达水平,探讨GNLY基因在肺结核病患者PBMC中的表达及其临床意义。方法应用已构建的质粒pGEM-T/GNLY作为定量模板行实时荧光定量RT-PCR测定收集自122例肺结核患者的标本,其中来自痰结核分枝杆菌阳性者62例(菌阳组),痰结核分枝杆菌阴性者60例(菌阴组)及40例健康对照者(对照组)外周血中GNLY mRNA的含量,并分析菌阳组抗结核治疗前后GNLY基因表达水平的变化。结果肺结核患者PBMC GNLY基因表达水平显著低于对照组(P<0.01);治疗前菌阳组PBMC GNLY基因表达水平显著低于菌阴组(P<0.05);治疗后菌阳组PBMC GNLY基因表达水平较治疗前显著升高(P<0.05)。结论本研究成功建立了检测人GNLY基因表达含量的荧光定量RT-PCR方法,GNLY基因表达水平可作为肺结核治疗的疗效观察及预后判断的一个新的实验室指标。Objective To establish a real-time quantitative reverse transcription polymerase chain reaction （RT-PCR） for detecting the expression of granulysin （GNLY） gene in peripheral blood mononuclear cells （PBMC） and explore the relationship between GNLY mRNA expression and pulmonary tuberculosis. Methods Real-time quantitative detection based on fluorescent TaqMan method was established. In this method, a constructed cloning vector pGEM-T/GNLY was taken as a standard plasmid. The GNLY gene expression levels in 122 patients with tuberculosis, including 62 patients with pulmonary tuberculosis of smear-positive, 60 of smear-negative and 40 healthy controls were detected by real-time RT-PCR. The expression changes of GNLY gene before and after treatment were also analyzed. Results The expression of GNLY gene in smear-positive tuberculosis group and smear-negative tuberculosis group were significantly lower than that in control group （P 0.01） and the mean level of GNLY mRNA in pulmonary tuberculosis of smear-positive group was significantly higher than that in smear-negative group （P0.05）, the expression of GNLY gene in smear-positive patients was significantly higher after treatment than before treatment （P 0.05）. Conclusions It is an easy and reliable test to detect the expression level of GNLY mRNA by real-time fluorescence quantitative RT-PCR and the preliminary experimental result shows that the detection of GNLY gene expression levels is meaningful for treatment guidance and evaluating prognosis of pulmonary tuberculosis.

关 键 词：颗粒溶素 结核 肺 逆转录聚合酶链反应 

分 类 号：R521[医药卫生—内科学] R440[医药卫生—临床医学]