title: Rapid molecular diagnosis of the Gilbert's syndrome-associated exon 1 mutation within the UGT1A1 gene.
abstract: Gilbert's syndrome is suspected in patients with unconjugated hyperbilirubinemia caused by decreased activity of the UDP-glucuronosyltransferase 1A1 (UGT1A1) gene in the absence of abnormal liver function and hemolysis. The major genetic variants underlying Gilbert's syndrome are TATA-box repeats of the promoter region and exon 1 G211A of the coding region, particularly in Asians. The efficacy of DNA melting curve analysis, however, has not been established for the G211A mutation. For rapid and accurate molecular diagnosis of Gilbert's syndrome, DNA melting curve analysis was evaluated for its genotyping capability not only for TATA-box repeats of the UGT1A1 promoter, but also for G211A of UGT1A1 exon 1. TA repeats within the TATA-box sequence and the exon 1 G211A mutation of the UGT1A1 gene were analyzed by DNA melting curve analysis. To evaluate the assay reliability, direct sequencing or polyacrylamide gel electrophoresis was used as a comparative method. All homozygous and heterozygous polymorphisms of A(TA)7TAA within the TATA-box allele and of exon 1 G211A mutants of the UGT1A1 gene were successfully identified with DNA melting curve analysis. DNA melting curve analysis is, therefore, an effective molecular method for the rapid diagnosis of Gilbert's syndrome, as it detects not only TATA-box polymorphisms but also the exon 1 G211A mutation located within the UGT1A1 gene.
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
authors.forename: T-Y
authors.initials: TY
authors.lastname: Hsieh
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
authors.forename: T-Y
authors.initials: TY
authors.lastname: Shiu
authors.affiliation: Department of Occupational Medicine, Taipei Medical University, Shuang-Ho Hospital, Taipei, Taiwan.
authors.forename: N-F
authors.initials: NF
authors.lastname: Chu
authors.affiliation: Division of Hematology and Oncology, Department of Internal Medicine, Taipei Medical University, Shuang-Ho Hospital, Taipei, Taiwan.
authors.forename: T-Y
authors.initials: TY
authors.lastname: Chao
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
authors.forename: H-C
authors.initials: HC
authors.lastname: Chu
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
authors.forename: W-K
authors.initials: WK
authors.lastname: Chang
authors.affiliation: Division of Gastroenterology, Buddhist Tzu Chi General Hospital, Taipei Branch, Taipei, Taiwan.
authors.forename: Y-C
authors.initials: YC
authors.lastname: Chao
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan xinhung@gmail.com.
authors.forename: H-H
authors.initials: HH
authors.lastname: Huang
mesh_terms: D000483:Alleles; D044466:Asian People / Q000235:genetics; D005091:Exons; D005838:Genotype; D005878:Gilbert Disease / Q000175:diagnosis / Q000235:genetics*; D014453:Glucuronosyltransferase / Q000235:genetics*; D006801:Humans; D009154:Mutation; D009691:Nucleic Acid Denaturation / Q000235:genetics; D057089:Pathology, Molecular; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D016385:TATA Box / Q000235:genetics; D000099148:UGT1A1 Enzyme
chemical_list: D014453:Glucuronosyltransferase; D000099148:UGT1A1 Enzyme