Laboratory of Clinical Molecular Biology, Institute of Biochemistry & Clinical Biochemistry, University Hospital A.Gemelli, Catholic University of the Sacred Heart, Rome, Italy.
Clin Chim Acta. 2010 Feb;411(3-4):246-9. doi: 10.1016/j.cca.2009.11.013. Epub 2009 Nov 20.
The basis of Gilbert's syndrome is a 70% reduction in bilirubin glucuronidation which, in the Caucasian population, is the result of a homozygous TA insertion into the promoter region of the UDP-glucuronosyltransferase 1A1 (UGT1A1) gene (UGT1A128 allele). In addition, homozygous subjects for UGT1A128 genotype may suffer from severe irinotecan toxicity or jaundice during treatment with the protease inhibitor atazanavir. For these reasons it is very important to perform a correct molecular diagnosis. In this study, we describe for the first time a new high resolution melting (HRM) analysis for a rapid UGT1A1 (TA)(n) genotyping.
We screened the TA number repetitions of the TATA-box promoter region of the UGT1A1 gene in 30 patients attending the Gemelli Hospital. In order to evaluate the reliability of this technique, we compared the results obtained by HRM and sequencing.
Since the TA insertion modifies the derivative melting curve shape and the melting temperature (T(m)), all possible genotypes for the 6 and 7 repeat alleles were successfully identified.
HRM analysis for the UGT1A1 (TA)(n) genotyping is a simple, rapid, sensitive and low cost method, very useful in diagnostics.
吉尔伯特综合征的基础是胆红素葡萄糖醛酸化减少 70%，在白种人群中，这是由于 UDP-葡萄糖醛酸基转移酶 1A1（UGT1A1）基因启动子区域的 TA 插入纯合子（UGT1A128 等位基因）所致。此外，UGT1A128 基因型的纯合子在接受蛋白酶抑制剂阿扎那韦治疗时可能会遭受严重的伊立替康毒性或黄疸。出于这些原因，进行正确的分子诊断非常重要。在这项研究中，我们首次描述了一种新的高分辨率熔解（HRM）分析方法，用于快速 UGT1A1（TA）（n）基因分型。
我们在 30 名就诊于 Gemelli 医院的患者中筛选了 UGT1A1 基因 TATA 盒启动子区域的 TA 重复数。为了评估该技术的可靠性，我们比较了 HRM 和测序的结果。
由于 TA 插入改变了衍生的熔解曲线形状和熔点（T(m)），因此可以成功鉴定出所有 6 重复和 7 重复等位基因的可能基因型。
用于 UGT1A1（TA）（n）基因分型的 HRM 分析是一种简单、快速、敏感且低成本的方法，在诊断中非常有用。
