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实时定量PCR检测隐球菌性脑膜炎中TRAIL表达
引用本文:周晔,;邓安梅,;陈燕,;谷明莉,;吴传勇,;陈波,;陈孙孝,;仲人前.实时定量PCR检测隐球菌性脑膜炎中TRAIL表达[J].解放军医学高等专科学校学报,2008(4):641-643.
作者姓名:周晔  ;邓安梅  ;陈燕  ;谷明莉  ;吴传勇  ;陈波  ;陈孙孝  ;仲人前
作者单位:[1]上海长征医院实验诊断科,上海200003
基金项目:本课题受国家自然科学基金(30671840、30772017)、上海市优秀学科带头人基金(07XD14013)资助.
摘    要:目的研究建立实时荧光定量(RFQ)-PCR法测定TRAIL mRNA含量的方法,探讨其在隐球菌性脑膜炎患者外周血单个核细胞中表达的检测价值,并分析隐球菌性脑膜炎患者免疫指标与TRAIL的相关性。方法设计特异性的引物和探针,以人基因GAPDH为内参照,用实时定量PCR的方法检测35例健康人和35例隐球菌性脑膜炎患者外周血单个核细胞中TRAIL mRNA的表达水平,采用特定蛋白分析仪测定IgG、IgA、IgM、C3、C4。结果与健康人比较,隐球菌性脑膜炎患者外周血单个核细胞中TRAIL mRNA的表达明显下降(P〈0.01)。隐脑患者血清中的C3和IgG显著下降(P〈0.01),且与TRAIL的变化成正相关。结论隐球菌性脑膜炎患者TRAIL下降,提示TRAIL可能参与隐球菌性脑膜炎的疾病进程,这为有效治疗隐球菌性脑膜炎提供了新的线索。

关 键 词:隐球菌性脑膜炎  TRAIL  PCR  实时定量

Correlation between TRAIL and Cryptococcal Meningitis
Institution:Zhou Ye,Deng An-mei,Chen Yan,Gu Ming-li,Wu Chuan-yong,Chen Bo,Chen Sun-xiao,Zhong Ren-qian (Department of Labortary Diagnostics,Shanghai Changzheng Hospital,Shanghai 200003,China)
Abstract:Objective To investigate the method of determining the content of TRAIL mRNA by real-time fluorescent quantitation (RFQ)-PCR, to explore the detection value of expression in peripheral blood mononuclear cells (PBMC) of patients with cryptococcal meningitis, and to analyze the correlation between immune indices of cryptococcal meningitis and TRAIL.Methods By designing specific primers and probes, and with human gene GAPDH as an internal reference, RFQ-PCR was employed to determine the levels of expression of TRAIL mRNA in PBMC from 35 healthy people and 35 patients with cryptococcal meningitis, and a single immunoquantitative diffusion method was adopted to determine the levels of IgG, IgA, IgM, C3, and C4. Results Compared with that of healthy people, the expression of TRAIL mRNA in PBMC of the patients with cryptococcal meningitis was significantly decreased (P〈0.01). The serum levels of C3 and IgG in the patients with cryptococcal meningitis were significantly decreased (P〈0.01), and were correlated positively with the change of TRAIL. Conclusion The significantly decreased expression of TRAIL mRNA indicates that TRAIL may be involved in the pathologic development of cryptococcal meningitis, thus providing a new approach for the effective treatment of cryptococcal meningitis.
Keywords:cryptococcal meningitis  TRAIL  polymerase chain reaction  real-time fluorescent quantitation
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