Rapid detection of the UGT1A1 single nucleotide polymorphism G211A using real-time PCR with Taqman Minor Groove Binder probes

Fei Liang Wong, May Kay Wang, Nem Yun Boo, Noor Hamidah Hussin, B. Othaman Ainoon

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The UGT1A1 Taqman MGB probe single nucleotide polymorphism (SNP) genotyping assay was developed to detect nucleotide 211 of the UDP-glucoronocyltransferase 1A1 (UGT1A1) gene. Defects in this enzyme interfere with process of conjugation of bilirubin and cause unconjugated hyperbilirubinemia. Variation at nucleotide 211 in the coding region of the UGT1A1 gene has been shown to be prevalent in Japanese and Chinese. Using an ABI sequence detection system (SDS) 7000, an allele-specific real-time PCR-based genotyping method was established to detect nucleotide G211A. Cord blood from 125 infants without hyperbilirubinemia (controls) were compared with cord blood from 74 infants (cases) with severe hyperbilirubinemia (total serum bilirubin >300 μmol/L). Homozygous variation of the UGT1A1 gene at nucleotide 211 (A/A) is significantly more common in cases (14.9%) than in controls (0.8%) (P<0.001). Direct sequencing from 20 randomly selected samples showed eight samples with homozygous wild type, seven with homozygous variant, and five samples were heterozygous. The result from this assay was in complete concordance with the DNA sequencing result and clearly discriminate wild-type (G/G), homozygous variant (A/A), and heterozygous (G/A). This assay is rapid and robust for screening of SNP G211A to determine if this polymorphism plays a role in causing severe neonatal jaundice in the local context.

Original languageEnglish
Pages (from-to)167-172
Number of pages6
JournalJournal of Clinical Laboratory Analysis
Volume21
Issue number3
DOIs
Publication statusPublished - 2007

Fingerprint

Uridine Diphosphate
Polymorphism
Hyperbilirubinemia
Binders
Single Nucleotide Polymorphism
Real-Time Polymerase Chain Reaction
Nucleotides
Fetal Blood
Bilirubin
Assays
Genes
Neonatal Jaundice
Blood
DNA Sequence Analysis
Alleles
Screening
Enzymes
Serum
Defects
DNA

Keywords

  • Hyperbilirubinemia
  • SNP
  • UGT1A1 gene

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Rapid detection of the UGT1A1 single nucleotide polymorphism G211A using real-time PCR with Taqman Minor Groove Binder probes. / Wong, Fei Liang; Wang, May Kay; Boo, Nem Yun; Hussin, Noor Hamidah; Othaman Ainoon, B.

In: Journal of Clinical Laboratory Analysis, Vol. 21, No. 3, 2007, p. 167-172.

Research output: Contribution to journalArticle

Wong, Fei Liang ; Wang, May Kay ; Boo, Nem Yun ; Hussin, Noor Hamidah ; Othaman Ainoon, B. / Rapid detection of the UGT1A1 single nucleotide polymorphism G211A using real-time PCR with Taqman Minor Groove Binder probes. In: Journal of Clinical Laboratory Analysis. 2007 ; Vol. 21, No. 3. pp. 167-172.
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