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Introducing a fast and simple PCR‐RFLP analysis for the detection of mutant thiopurine S ‐methyltransferase alleles TPMT*3A and TPMT*3C
Author(s) -
Oender K,
Lanschuetzer CM,
Laimer M,
Klausegger A,
Paulweber B,
Kofler B,
Hintner H,
Bauer JW
Publication year - 2006
Publication title -
journal of the european academy of dermatology and venereology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.655
H-Index - 107
eISSN - 1468-3083
pISSN - 0926-9959
DOI - 10.1111/j.1468-3083.2006.01459.x
Subject(s) - thiopurine methyltransferase , azathioprine , medicine , restriction fragment length polymorphism , allele , genotyping , polymerase chain reaction , pharmacology , genetics , biology , genotype , gene , disease
Background Azathioprine, in combination with corticosteroids, is the first‐line therapy of severe forms of pemphigus vulgaris. Patients with an impaired thiopurine S ‐methyltransferase (TPMT) activity are at risk of developing severe myelo‐suppression upon treatment with thiopurines such as azathioprine. Analysis of the TPMT status prior to drug administration is therefore highly recommended. However, because of the limited availability of TPMT testing outside of specialized centres, pre‐emptive TPMT testing is not widespread. To avoid laborious biochemical and sequencing assays, we evaluated a new restriction fragment length polymorphism (RFLP) analysis. Methods We designed a rapid genetic polymerase chain reaction (PCR)‐RFLP screen for the most prevalent mutant TPMT*3A and TPMT*3C alleles that are known to result in reduced TPMT enzyme activity. Results Validating our fast system on 871 Caucasian DNA samples, we observed that 8.61% of our probands carried the TPMT*3A allele and 0.23% were heterozygous for the TPMT*3C allele, which is in concordance with previously reported allele frequencies. Conclusion This simple and low‐cost PCR‐RFLP TPMT polymorphism testing approach can be performed in a standard laboratory. It should be applied to all patients prior to receiving thiopurine drug therapy to avoid the severe, but predictable, haematopoietic side‐effects of thiopurine drug administration.