运用TaqMan探针实时荧光PCR技术检测AKAP10基因2073A/G单核苷酸多态性  

Genotyping of AKAP10 Gene 2073A/G Single Nucleotide Polymorphism by TaqMan Probe Real-time PCR

作　　者：王默进[1] 周总光[1] 王玲[1] 李园[1] 张鹏[1] 张益[1] 崔长富[1] 周斌[1] 

机构地区：[1]四川大学华西医院消化外科研究室,成都610041

出　　处：《四川大学学报（医学版）》2009年第2期275-278,共4页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号3057181);美国中华医学基金(No.CMB96636)资助

摘　　要：目的建立一套以TaqMan探针实时荧光PCR技术为基础的单核苷酸多态性（SNP）检测平台，并用于AKAPIO基因2073A／GSNP的检测。方法设计一对TaqMan探针加入到PCR反应系统中，并设立阴阳对照，对AKAPIO基因2073A／GSNP进行分型。同时验证部分样本PCR产物的直接测序结果。结果运用TaqMan探针实时荧光PCR技术对AKAPIO基因2073A／GSNP检测结果准确，与PCR产物的直接测序结果完全一致。AKAPIO基因2073A／G多态性分布与性别年龄无关（P〉0．05）。非条件logistic回归分析提示，AKAP10基因2073A／G突变型（AG＋GG）与野生型AA相比增加结直肠癌的发生风险（OR=1．52，95％CI：1．07～2．15。P=0．019）。结论通过合理设计TaqMan探针，设立阴阳性对照，应用TaqMan探针实时荧光PCR技术成功建立起一套SNP检测平台。AKAP10基因2073A／G多态性与结直肠癌发病存在显著相关性。Objective To detect AKAPIO gene 2073A/G single nucleotide polymorphism （SNP） genotyping by TaqMan probe real-time PCR. Methods The genotype of AKAPIO gene 2073A/G was detected by real-time PCR with a pair of new-designed TaqMan probes. The PCR products also were subjected to gene sequence analysis to validate the results of TaqMan probe real-time PCR. Results The TaqMan probe real-time PCR method was successfully developed to detect AKAPIO gene 2073A/G SNP. The results were accordant with those achieved by DNA sequencing. The distribution of AKAPIO gene 2073A/G among population had no relationship with gender, age （P〉0.05）. Unconditional logistic regression analysis revealed that the variant genotypes （AG ＋ GG） had a 52% increased risk of colorectal cancer, compared with the AA genotype （P=0. 019）. Conclusion A detection platform for SNP genotyping by TaqMan probe was set up successfully. There was a significant association between AKAPIO gene 2073A/G and colorectal cancer.

关 键 词：TAQMAN探针 实时荧光PCR 单核苷酸多态性 

分 类 号：R450[医药卫生—治疗学]