Department of Public Health and Pediatrics, School of Medicine, University of Turin, Piazza Polonia 94, 10126, Turin, Italy.
BioMole srl., Via Quarello 15/A, 10100, Turin, Italy.
Mol Biol Rep. 2021 May;48(5):4953-4959. doi: 10.1007/s11033-021-06454-2. Epub 2021 Jun 4.
Gilbert's syndrome is characterized by mild unconjugated hyperbilirubinemia. The key of this disease is a diminished activity of UDP-glucuronosyltransferase 1A1 (UGT1A1). TA insertion into the TATA box promoter region of the UGT1A1 gene on chromosome 2 is the genetic basis of Gilbert's syndrome (UGT1A128). An extra TA insert leads to eight (TA)8 repeats (UGT1A137) resulting in a further reduction of glucuronidation activity. A variant lacking one TA repeat (TA)5 (UGT1A136) has been identified. (TA)8 repeats (UGT1A137) and (TA)5 (UGT1A136) have been detected in Africans (frequency up to 0.07 and 0.08 respectively). We present a real time PCR method for genotyping the UGT1A1 (TA)n polymorphism (UGT1A128, UGT1A136, UGT1A137) using Taqman PCR on 7500 and cfx96 Real-Time PCR System. We present a real time PCR method for genotyping the UGT1A1 (TA)n polymorphism (UGT1A128, UGT1A136, UGT1A1*37) using Taqman PCR. About clinical validation, all 53 samples collected from patients referred for suspected Gilbert's syndrome were analyzed. We found 21 on the 53 patients (39.6%) were homozygotes (UGT1A1-TATA (TA)6) and referred as wild-type, 13 on the 53 patients (24.5%) were homozygotes (UGT1A1-TATA (TA)7) and referred as mutated, 1 on the 53 patients (1.9%) were homozygotes (UGT1A1-TATA (TA)8) and referred as mutated, 1 on the 53 patients (1.9%) were heterozygotes (UGT1A1-TATA (TA)7/8) and referred as mutated, 1 on the 53 patients (1.9%) were heterozygotes (UGT1A1-TATA (TA)5/6) and referred as mutated, and 16 on the 53 patients (30.2%) were heterozygotes (UGT1A1-TATA (TA)6/7). None were homozygotes UGT1A1-TATA (TA)5, homozygotes UGT1A1-TATA (TA)8, or heterozygotes with (TA)5 or (TA)8 alleles. The newly described technique represents a valid alternative method to sequencing, mainly due to its rapidity, easiness, and minor costs.
吉尔伯特综合征的特征是轻度非结合性高胆红素血症。这种疾病的关键是 UDP-葡糖醛酸基转移酶 1A1（UGT1A1）活性降低。TA 插入 UGT1A1 基因 TATA 盒启动子区在染色体 2 上是吉尔伯特综合征（UGT1A128）的遗传基础。额外的 TA 插入导致 8 个（TA）8 重复（UGT1A137），导致葡萄糖醛酸化活性进一步降低。已经鉴定出一种缺少一个 TA 重复（TA）5（UGT1A136）的变体。在非洲人（频率分别高达 0.07 和 0.08）中检测到（TA）8 重复（UGT1A137）和（TA）5（UGT1A136）。我们提出了一种使用 Taqman PCR 在 7500 和 cfx96 实时 PCR 系统上对 UGT1A1（TA）n 多态性（UGT1A128、UGT1A136、UGT1A137）进行基因分型的实时 PCR 方法。我们提出了一种使用 Taqman PCR 对 UGT1A1（TA）n 多态性（UGT1A128、UGT1A136、UGT1A1*37）进行基因分型的实时 PCR 方法。关于临床验证，我们分析了从疑似吉尔伯特综合征患者中收集的 53 个样本。我们发现 53 名患者中的 21 名（39.6%）为纯合子（UGT1A1-TATA（TA）6），并被归为野生型，53 名患者中的 13 名（24.5%）为纯合子（UGT1A1-TATA（TA）7），并被归为突变型，53 名患者中的 1 名（1.9%）为纯合子（UGT1A1-TATA（TA）8），并被归为突变型，53 名患者中的 1 名（1.9%）为杂合子（UGT1A1-TATA（TA）7/8），并被归为突变型，53 名患者中的 1 名（1.9%）为杂合子（UGT1A1-TATA（TA）5/6），并被归为突变型，53 名患者中的 16 名（30.2%）为杂合子（UGT1A1-TATA（TA）6/7）。没有一个是 UGT1A1-TATA（TA）5 纯合子，UGT1A1-TATA（TA）8 纯合子，或（TA）5 或（TA）8 等位基因的杂合子。新描述的技术代表了一种替代测序的有效方法，主要是由于其快速性、易用性和较低的成本。
