荧光定量PCR溶解曲线分析技术检测多系统萎缩患者TCRβ链CDR3谱序初探  

The measurment of the CDR3 gene spectratype of TCR beta chain by FQ-PCR melting cure in mutiple system atrophy

作　　者：马锐[1] 周建伟[1] 胡雅丽[2] 姚新生[1] 

机构地区：[1]遵义医学院免疫学教研室,贵州遵义563003 [2]遵义医学院第三附属医院,贵州遵义563000

出　　处：《遵义医学院学报》2009年第1期1-4,共4页Journal of Zunyi Medical University

基　　金：国家重点基础研究发展规划973前期研究专项(2008CB517310);国家自然科学基金(30760231);教育部科学技术研究项目9208127);贵州省科技厅项目(2008GZ00516&20072122)资助

摘　　要：目的利用荧光定量PCR溶解曲线分析技术检测多系统萎缩病人TCRβ链CDR3谱序。方法提取1例多系统萎缩病人外周血单个核细胞中的总RNA逆转录成cDNA,以26个人TRBV基因家族设计上游引物,共同的TRBC基因设计下游引物,用荧光定量PCR(FQ-PCR)扩增26个TRBV基因各家族CDR3谱系,根据其溶解曲线图分析各家族CDR3谱系的单/寡/多克隆增生。结果该病例外周血T细胞TCRβ链26个家族CDR3表达频率不一致,有的家族呈现缺失状态,有的家族出现寡克隆状态。结论该多系统萎缩病人TCRβ链CDR3存在谱系漂移,有的表达呈寡克隆,有的无表达呈缺失。Objective To explore the CDR3 gene spectratype of TCR beta chain in a patient with multiple system atrophy using real-time fluorescence quantitative reverse transcription polymerase chain reaction （FQ-PCR） with DNA melting curve analysis. Methods The total RNA of peripheral blood mononuelear cells （PBMCs） from patient with multiple system atrophy was transcripted reversely into cDNA,designed the primer of TRBV and TRBC,amplied cDNA by FQ-PCR,and analyzed the state of monoclonal/oligoclonal/polyclonal CDR3 gene. Results The FQ-PCR products of 26 TRBV families CDR3 showed different frequency ,parts of TRBV families CDR3 products disappeared and parts showed single peaks. Conclusion Compared to healthy control,the CDR3 gene of TCR BV showed monoclonal or vicious.

关 键 词：多系统萎缩 互补决定区 荧光定量PCR 溶解曲线 

分 类 号：R392[医药卫生—免疫学]