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Rapid genotyping of common deficient thiopurine S-methyltransferase alleles using the DNA-microchip technique

Rapid genotyping of common deficient thiopurine S-methyltransferase alleles using the... Thiopurine drugs are metabolized, in part, by S-methylation catalyzed by thiopurine S-methyltransferase (TPMT). Patients with very low or undetectable TPMT activity are at high risk of severe, potentially fatal hematopoietic toxicity when they are treated with standard doses of thiopurines. As human TPMT activity is controlled by a common genetic polymorphism, it is an excellent candidate for the clinical application of pharmacogenetics. Here, we report a new molecular approach developed to detect point mutations in the TPMT gene that cause the loss of TPMT activity. A fluorescently labeled amplified DNA is hybridized with oligonucleotide DNA probes immobilized in gel pads on a biochip. The specially designed TPMT biochip can recognize six point mutations in the TPMT gene and seven corresponding alleles associated with TPMT deficiency: TPMT * 2; TPMT * 3A, TPMT * 3B, TPMT * 3C, TPMT * 3D, TPMT * 7, and TPMT * 8. The effectiveness of the protocol was tested by genotyping 58 samples of known genotype. The results showed 100% concordance between the biochip-based approach and the established PCR protocol. The genotyping procedure is fast, reliable and can be used for rapid screening of inactivating mutations in the TPMT gene. The study also provides the first data on the frequency of common TPMT variant alleles in the Russian population, based on a biochip analysis of 700 samples. TPMT gene mutations were identified in 44 subjects; genotype *1/*3A was most frequent. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png European Journal of Human Genetics Springer Journals

Rapid genotyping of common deficient thiopurine S-methyltransferase alleles using the DNA-microchip technique

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References (30)

Publisher
Springer Journals
Copyright
Copyright © 2006 by Nature Publishing Group
Subject
Biomedicine; Biomedicine, general; Human Genetics; Bioinformatics; Gene Expression; Cytogenetics
ISSN
1018-4813
eISSN
1476-5438
DOI
10.1038/sj.ejhg.5201647
Publisher site
See Article on Publisher Site

Abstract

Thiopurine drugs are metabolized, in part, by S-methylation catalyzed by thiopurine S-methyltransferase (TPMT). Patients with very low or undetectable TPMT activity are at high risk of severe, potentially fatal hematopoietic toxicity when they are treated with standard doses of thiopurines. As human TPMT activity is controlled by a common genetic polymorphism, it is an excellent candidate for the clinical application of pharmacogenetics. Here, we report a new molecular approach developed to detect point mutations in the TPMT gene that cause the loss of TPMT activity. A fluorescently labeled amplified DNA is hybridized with oligonucleotide DNA probes immobilized in gel pads on a biochip. The specially designed TPMT biochip can recognize six point mutations in the TPMT gene and seven corresponding alleles associated with TPMT deficiency: TPMT * 2; TPMT * 3A, TPMT * 3B, TPMT * 3C, TPMT * 3D, TPMT * 7, and TPMT * 8. The effectiveness of the protocol was tested by genotyping 58 samples of known genotype. The results showed 100% concordance between the biochip-based approach and the established PCR protocol. The genotyping procedure is fast, reliable and can be used for rapid screening of inactivating mutations in the TPMT gene. The study also provides the first data on the frequency of common TPMT variant alleles in the Russian population, based on a biochip analysis of 700 samples. TPMT gene mutations were identified in 44 subjects; genotype *1/*3A was most frequent.

Journal

European Journal of Human Genetics – Springer Journals

Published: May 17, 2006

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