Startseite Microarray with LNA-probes for genotyping of polymorphic variants of Gilbert’s syndrome gene UGT1A1(TA)n
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Microarray with LNA-probes for genotyping of polymorphic variants of Gilbert’s syndrome gene UGT1A1(TA)n

  • Eugeny E. Fesenko EMAIL logo , Rustam N. Heydarov , Eugenia V. Stepanova , Michael E. Abramov , Alexander V. Chudinov , Alexander S. Zasedatelev und Vladimir M. Mikhailovich
Veröffentlicht/Copyright: 15. Dezember 2012
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Clinical Chemistry and Laboratory Medicine (CCLM)
Aus der Zeitschrift Clinical Chemistry and Laboratory Medicine (CCLM) Band 51 Heft 6

Abstract

Background: Gilbert’s syndrome is a common metabolic dysfunction characterized by elevated levels of unconjugated bilirubin in the bloodstream. This condition is usually caused by additional (TA) insertions in a promoter region of the uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1) gene, which instead of the sequence А(TА)6TАА contains А(TА)7TАА. While the condition itself is benign, it presents elevated risk for patients treated with irinotecan, a common chemotherapy drug.

Methods: The technique is based on hybridization analysis of a pre-amplified segment of the UGT1A1 gene promoter performed on a microarray. Specific probes containing locked nucleic acids (LNA) were designed and immobilized on the microarray to provide accurate identification.

Results: A microarray has been developed to identify both common and rare variants of UGT1A1(TA)n polymorphisms. In total, 108 individuals were genotyped. Out of these, 47 (43.5%) had homozygous wild-type genotypes (TA)6/(TA)6; 41(38%) were heterozygotes (TA)6/(TA)7; and 18 (16.7%) – homozygotes (TA)7/(TA)7. In two cases (1.8%), rare genotypes (TA)5/(TA)7and (TA)5/(TA)6were found. The results were in full agreement with the sequencing. In addition, synthetic fragments corresponding to all human allelic variants [(TA)5, (TA)6, (TA)7, (TA)8] were successfully tested.

Conclusions: The developed microarray-based approach for identification of polymorphic variants of the UGT1A1 gene is a promising and reliable diagnostic tool that can be successfully implemented in clinical practice.

Keywords: genotyping; locked nucleic acids; microarray; TA-repeats; UGT1A1; uridine diphosphate glucuronosyltransferase
Corresponding author: Eugeny E. Fesenko, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Street 32, Moscow 119991, Russia

This work was partially supported by contract No. 16.512.11.2042 with the Federal Agency of Sciences and Innovations of the Russian Federation. We are grateful to E. Chernoglazova for clinical data, to E. Kreindlin for manufacturing the microarrays, to S. Surzhikov and O. Ivashkina for the synthesis of oligonucleotides and to V. Chechetkin for his help with the mathematical calculations and analysis of experimental data. We are especially grateful to A. Kolchinsky for his assistance in the preparation of this paper.

Conflict of interest statement

Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article.

Research funding: Partially supported by contract No. 16.512.11.2042 with the Federal Agency of Sciences and Innovations of the Russian Federation.

Employment or leadership: None declared.

Honorarium: None declared.

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Received: 2012-09-29
Accepted: 2012-11-09
Published Online: 2012-12-15
Published in Print: 2013-06-01

©2013 by Walter de Gruyter Berlin Boston

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https://doi.org/10.1515/cclm-2012-0656
Schlagwörter für diesen Artikel
genotyping; locked nucleic acids; microarray; TA-repeats; UGT1A1; uridine diphosphate glucuronosyltransferase

Artikel in diesem Heft

  1. Letters to the Editor
  2. Evaluation of a novel room temperature RNA storage tube for use in a real-time quantitative PCR assay
  3. Evaluation of the Universal Master Mix (STAT-NAT DNA-Mix) for reliable molecular testing
  4. HbA1c: performance of the Sebia Capillarys 2 Flex Piercing
  5. A case of monoclonal gammopathy of undetermined significance (MGUS): type IgD-lambda
  6. Evidence-based use of serum protein electrophoresis in laboratory medicine
  7. Plasma volume shifts during multiday racing
  8. Structured handoff at shift change in a clinical laboratory increases patient safety
  9. Instrument-dependent interference of Howell-Jolly bodies in reticulocyte enumeration
  10. Reply to Gore et al.: Plasma volume shift during multiday racing
  11. Recommendations for appropriate serum electrophoresis requests: the Italian approach
  12. Masthead
  13. Masthead
  14. Editorials
  15. Biomarkers for sepsis: an unfinished journey
  16. Laboratory demand management of repetitive testing – time for harmonisation and an evidenced based approach
  17. Reviews
  18. Non-invasive prenatal diagnostics of aneuploidy using next-generation DNA sequencing technologies, and clinical considerations
  19. The diagnostic utility of brain natriuretic peptide in heart failure patients presenting with acute dyspnea: a meta-analysis
  20. Opinion Paper
  21. Opinion paper on innovative approach of biomarkers for infectious diseases and sepsis management in the emergency department
  22. Genetics and Molecular Diagnostics
  23. Microarray with LNA-probes for genotyping of polymorphic variants of Gilbert’s syndrome gene UGT1A1(TA)n
  24. Selection of the optimal manual method of cell free fetal DNA isolation from maternal plasma
  25. A multiplex assay to rapidly exclude HLA-DQ2.5 and HLA-DQ8 expression in patients at risk for celiac disease
  26. Low cost biosensor-based molecular differential diagnosis of α-thalassemia (Southeast Asia deletion)
  27. General Clinical Chemistry and Laboratory Medicine
  28. Managing laboratory test ordering through test frequency filtering
  29. Critical review of laboratory investigations in clinical practice guidelines: proposals for the description of investigation
  30. Long-term stability of laboratory tests and practical implications for quality management
  31. Coffee consumption, serum γ-glutamyltransferase, and glucose tolerance status in middle-aged Japanese men
  32. Biological variation and prognosis usefulness of new biomarkers in liver transplantation
  33. Switching between parathormone (PTH) assays: the impact on the diagnosis of renal osteodystrophy
  34. A comparison between two different in vitro basophil activation tests for gluten- and cow’s milk protein sensitivity in irritable bowel syndrome (IBS)-like patients
  35. A proficiency testing program of hemoglobin analysis in prevention and control of severe hemoglobinopathies in Thailand
  36. Advancing haemostasis automation – successful implementation of robotic centrifugation and sample processing in a tertiary service hospital
  37. ADAM12s and PP13 as first trimester screening markers for adverse pregnancy outcome
  38. Analysis of serous body fluids using the CELL-DYN Sapphire hematology analyzer
  39. Cancer Diagnostics
  40. Association and prognostic value of serum inflammation markers in patients with leukoplakia and oral cavity cancer
  41. Do laboratories follow heart failure recommendations and guidelines and did we improve? The CARdiac MArker Guideline Uptake in Europe (CARMAGUE)
  42. Cardiovascular Diseases
  43. Additional diagnostic and prognostic value of copeptin ultra-sensitive for diagnosis of non-ST-elevation myocardial infarction in older patients presenting to the emergency department1)
  44. Moderate elevations of high-sensitivity cardiac troponin I and B-type natriuretic peptide in chronic hemodialysis patients are associated with mortality
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