荧光偏振技术分析全血XPD基因单核苷酸多态性的研究  

Detection of Single Nuclear Polymorphisms of Xeroderma Pigmentosum Group D in Whole Blood by Fluorescence Polarization Assay

作　　者：李丁[1] 张菊[1] 颜真[1] 张贺龙[2] 郭宴海[1] 刘文超[3] 李小惠[2] 李宏伟[1] 

机构地区：[1]第四军医大学国家肿瘤生物学重点实验室,药学系全军基因芯片重点实验室,全军基因诊断技术研究所,西安710033 [2]第四军医大学唐都医院肿瘤科,西安710038 [3]第四军医大学西京医院肿瘤科,西安710032

出　　处：《分析化学》2010年第4期585-588,共4页Chinese Journal of Analytical Chemistry

基　　金：国家高技术研究和发展计划基金(No.2008AA02Z444)资助项目

摘　　要：将荧光偏振与非对称基因扩增技术联用,建立了可用于检测全血XPD基因单核苷酸多态性的新方法。用不等量(1∶5)的XPD基因上、下游引物对含单核苷酸多态性位点的目的片段进行非对称扩增,再用两种单核苷酸多态性序列特异的荧光标记探针对扩增产物进行检测。由于扩增得到的单链片段能够与各自不同的荧光标记探针特异结合,使荧光标记分子的分子量增加,偏振值(FP)增高。通过检测增高的FP值,可确定目的片段单核苷酸多态性。采用本方法对98例全血的XPD基因第751位密码子进行了单核苷酸多态性分析,并与传统的荧光偏振检测方法进行了比较,取得满意结果。A method for the detection of xeroderma pigmentosum group D gene（XPD）polymorphisms was developed based on the fluorescence polarization and asymmetric PCR assay.First,the target gene was amplified in asymmetric PCR by unequal amount of primers,the single-stranded amplicons in PCR product were identified by hybridization.The hybridization between the fluroscence-labeled probes and the single-stranded amplicons resulted in the increase of the fluorescence polarization values.XPD polymorphisms were identified by the increased fluorescence polarization values and 98 samples were analyzed by template-directed dye-terminator incorporation with fluorescence polarization（TDI-FP）and the fluorescence polarization-asymmetric PCR assay in parallel.This study demonstrated that the fluorescence polarization-asymmetric PCR assay was a simple,specific,sensitive and cost-effective method for the detection of single nucleotide polymorphisms.

关 键 词：单核苷酸多态性 XPD基因 荧光偏振检测 

分 类 号：R450[医药卫生—治疗学]