Analysis on A(TA)n TAA polymorphism of UGT1A1 gene promoter by fluorescence real-time quantitative PCR
10.3969/j.issn.1673-4130.2016.13.023
- VernacularTitle:应用实时荧光定量PCR技术分析 UGT1A1基因启动子区 A（TA）nTAA 多态性
- Author:
Yuzhong XU
;
Qunrong CHEN
;
Shunchang SUN
;
Yunsheng PENG
- Publication Type:Journal Article
- Keywords:
UGT1A1 gene;
polymorphism;
fluorescence real-time quantitative PCR
- From:
International Journal of Laboratory Medicine
2016;37(13):1806-1808
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a new method to detect A (TA)n TAA polymorphism in the UGT1A1 gene promoter by fluo‐rescence real‐time quantitative PCR (RQ‐PCR) .Methods Genomic DNA was extracted from peripheral blood in 16 patients with Gilbert′s syndrome and 66 healthy individuals .The polymorphic A(TA)n TAA sequence in the UGT1A1 gene promoter was deter‐mined by DNA sequencing .A pair of primers and two TaqMan probes labeled with either 5′FAM or VIC reporter dye incorporated a 3′minor groove binder were designed .The A(TA)n TAA polymorphisms in the UGT1A1 gene promoter were identified by RQ‐PCR for all research subjects .The sensitivity and specificity of RQ‐PCR for detecting the A(TA)nTAA polymorphisms were veri‐fied by DNA sequencing method .Results The homozygous A(TA)7TAA polymorphism was found in the promoter region of the UGT1A1 gene in all 16 patients with Gilbert′s syndrome by using RQ‐PCR .The homozygous A(TA)6TAA polymorphism was foundin46healthysubjects,whiletheheterozygousA(TA)6TAA/A(TA)7TAApolymorphismwasfoundinother20healthysub‐jects .All A(TA)nTAA polymorphisms in the promoter region of the UGT1A1 gene identified by RQ‐PCR were consistent with that of DNA sequencing .Conclusion It is a sensitive ,specific and simple method to detect the A (TA)n TAA polymorphisms in the promoter region of the UGT1A1 gene by RQ‐PCR ,which can be promoted and applied in clinic .
