Tsai Shin-Yi, Er Tze-Kiong, Lin Chin-Wen, Chang Jan-Gowth
Clin Lab. 2014;60(6):1015-26. doi: 10.7754/clin.lab.2013.130707.
Uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1) is a key conjugating enzyme of bilirubin and the anti-tumor medication irinotecan. Comprehensive analysis of UGT1A1 gene polymorphisms may provide benefit by predicting pharmacokinetics and outcomes of treatment with irinotecan and certain antiviral medications.
A high-resolution melting (HRM) analysis was designed to characterize the UGT1A1 gene. Genomic DNA from 110 healthy subjects was extracted from peripheral blood samples. The promoter and 11 exons from UGT1A1 were screened by HRM, and all results were subsequently confirmed by direct DNA sequencing.
HRM analysis readily identified UGT1A1 gene mutations. We identified 5 different known variants of UGT1A1 including c.211 G > A; G71R, c.686 C > A; P229Q, c.1091 C > T; c.-3279 T > G; and c.-3156 G > A in 110 normal Taiwanese individuals. We also identified 8 new sequence variants, namely, c.-3296 C > T; c.43 C > A; c.45 G > A; c.234 G > A; c.577 G > A; c.614 C > T; c.1011 T > C; and c.1352 C > T. Each UGT1A1 variant was easily identifiable by differences in curves plotted from HRM data.
HRM analysis was rapid, accurate, and economical for screening UGT1A1 gene mutations.
尿苷二磷酸葡萄糖醛酸基转移酶1A1（UGT1A1）是胆红素和抗肿瘤药物伊立替康的关键结合酶。对UGT1A1基因多态性进行综合分析，可能有助于预测伊立替康及某些抗病毒药物的药代动力学和治疗效果。
设计了一项高分辨率熔解（HRM）分析来鉴定UGT1A1基因。从110名健康受试者的外周血样本中提取基因组DNA。通过HRM对UGT1A1的启动子和11个外显子进行筛查，随后所有结果均通过直接DNA测序进行确认。
HRM分析能够轻松鉴定出UGT1A1基因突变。我们在110名台湾正常个体中鉴定出5种不同的已知UGT1A1变体，包括c.211 G>A；G71R、c.686 C>A；P229Q、c.1091 C>T、c.-3279 T>G和c.-3156 G>A。我们还鉴定出8种新的序列变体，即c.-3296 C>T、c.43 C>A、c.45 G>A、c.234 G>A、c.577 G>A、c.614 C>T、c.1011 T>C和c.1352 C>T。通过HRM数据绘制的曲线差异，可轻松识别每种UGT1A1变体。
HRM分析在筛查UGT1A1基因突变方面快速、准确且经济。
