一步法鉴定UGT1A1基因启动子中的所有长度多态性。

Single-step identification of all length polymorphisms in the UGT1A1 gene promoter.

作者信息

Skarke C, Grösch S, Geisslinger G, Lötsch J

机构信息

Pharmazentrum Frankfurt, Institute of Clinical Pharmacology, Johann Wolfgang Goethe University, Frankfurt, Germany.

出版信息

Int J Clin Pharmacol Ther. 2004 Mar;42(3):133-8. doi: 10.5414/cpp42133.

DOI:10.5414/cpp42133

PMID:15049432

Abstract

AIM

To provide a sensitive genetic screening method for rapid identification of all known length polymorphisms in the promoter region of the uridine 5'-diphosphoglucose glucuronosyltransferase (UGT) 1A1 gene comprising (TA)5, (TA)7 and (TA)8 repeats as opposed to the non-mutated (TA)6 allele.

METHODS

The UGT1A1 promoter genotype was assessed in 115 subjects by means of a newly developed pyrosequencing method. PCR-generated DNA templates of heterozygous (TA)5 and (TA)7 carriers were cloned into a TOPO TA vector and verified by sequencing. In addition, a (TA)8 segment was produced by cloning to demonstrate the ability of the method to detect this mutation.

RESULTS

All length polymorphisms of the UGT1A1 promoter described in the literature were clearly identified. Fifteen subjects had Gilbert's syndrome with elevated serum bilirubin associated with a homozygous (TA)7TAA/(TA)7TAA genotype. Two subjects with the rare genotypes (TA)5TAA/(TA)6TAA and (TA)5TAA/(TA)7TAA were found, where only the latter one displayed elevated serum bilirubin levels. Allelic frequencies were 0.9%, 66.1% and 33% for the (TA)5TAA, (TA)6TAA and (TA)7TAA allele, respectively.

CONCLUSION

Our method enables reliable genetic single-step screening for all known length polymorphisms in the UGT1A1 gene promoter that cause Gilbert's syndrome. This facilitates pharmacogenetic-guided dosing of drugs with known toxicity metabolized by UGT1A1.

摘要

目的

提供一种灵敏的基因筛查方法，用于快速鉴定尿苷二磷酸葡萄糖醛酸基转移酶（UGT）1A1基因启动子区域所有已知的长度多态性，包括（TA）5、（TA）7和（TA）8重复序列，以区别于未突变的（TA）6等位基因。

方法

采用新开发的焦磷酸测序法对115名受试者的UGT1A1启动子基因型进行评估。将杂合（TA）5和（TA）7携带者的PCR扩增DNA模板克隆到TOPO TA载体中，并通过测序进行验证。此外，通过克隆产生一个（TA）8片段，以证明该方法检测此突变的能力。

结果

文献中描述的UGT1A1启动子的所有长度多态性均被清晰鉴定。15名受试者患有吉尔伯特综合征，血清胆红素升高，与纯合（TA）7TAA/（TA）7TAA基因型相关。发现两名具有罕见基因型（TA）5TAA/（TA）6TAA和（TA）5TAA/（TA）7TAA的受试者，其中只有后者血清胆红素水平升高。（TA）5TAA、（TA）6TAA和（TA）7TAA等位基因的频率分别为0.9%、66.1%和33%。