title: Three-dimensional polyacrylamide gel-based DNA microarray method effectively identifies UDP-glucuronosyltransferase 1A1 gene polymorphisms for the correct diagnosis of Gilbert's syndrome.
abstract: Gilbert's syndrome is a mild genetic liver disorder characterized by unconjugated hyperbilirubinemia due to defects in the UDP-glucuronosyltransferase 1A1 (UGT1A1) gene. The T-3279G mutation in the phenobarbital responsive enhancer module (PBREM), the TA-insertion in the TATA box, creating the A(TA)7TAA motif instead of A(TA)6TAA and the G211A mutation in coding exon 1, particularly in Asian populations, of the human UGT1A1 gene are the three common genotypes found in patients with Gilbert's syndrome. Different approaches for detecting the T-3279G, A(TA)6/7TAA and G211A mutations of the UGT1A1 gene have been described. In this study, to the best of our knowledge, we established a three-dimensional polyacrylamide gel-based DNA microarray method for the first time, in order to study UGT1A1 gene polymorphisms. This method, based on a step-by-step three-dimensional polyacrylamide gel-based DNA microarray protocol, successfully identified all possible genotypes of T-3279G, A(TA)6/7TAA and G211A in 20 patients with hyperbilirubinemia. In addition, sequencing was performed to confirm these results. The data from the current study demonstrate that the three-dimensional polyacrylamide gel microarray method has the potential to be applied as a useful, reliable and cost-effective tool to detect the T-3279G, the A(TA)6/7TAA and the G211A mutations of the UGT1A1 gene in patients with hyperbilirubinemia and thereby aid in the diagnosis of Gilbert's syndrome.
authors.affiliation: The Second Hospital of Nanjing, Affiliated to the Medical School of Southeast University, Nanjing, Jiangsu 210003, P.R. China.
authors.forename: Jinyun
authors.initials: J
authors.lastname: Song
authors.affiliation: The Second Hospital of Nanjing, Affiliated to the Medical School of Southeast University, Nanjing, Jiangsu 210003, P.R. China.
authors.forename: Mei
authors.initials: M
authors.lastname: Sun
authors.affiliation: The Second Hospital of Nanjing, Affiliated to the Medical School of Southeast University, Nanjing, Jiangsu 210003, P.R. China.
authors.forename: Jiayan
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Child Development and Learning Science, Southeast University, Nanjing, Jiangsu 210096, P.R. China.
authors.forename: Dongrui
authors.initials: D
authors.lastname: Zhou
authors.affiliation: The Second Hospital of Nanjing, Affiliated to the Medical School of Southeast University, Nanjing, Jiangsu 210003, P.R. China.
authors.forename: Xuping
authors.initials: X
authors.lastname: Wu
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry*; D044466:Asian People; D005260:Female; D005838:Genotype; D005878:Gilbert Disease; D014453:Glucuronosyltransferase / Q000235:genetics*; D006801:Humans; D008297:Male; D009154:Mutation / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011110:Polymorphism, Genetic / Q000235:genetics*; D000099148:UGT1A1 Enzyme
chemical_list: D000180:Acrylic Resins; D014453:Glucuronosyltransferase; D000099148:UGT1A1 Enzyme; C016680:polyacrylamide gels