title: Introducing a fast and simple PCR-RFLP analysis for the detection of mutant thiopurine S-methyltransferase alleles TPMT*3A and TPMT*3C.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Dermatology, Paracelsus Private Medical University, Salzburg, Austria.
authors.forename: K
authors.initials: K
authors.lastname: Oender
authors.forename: C M
authors.initials: CM
authors.lastname: Lanschuetzer
authors.forename: M
authors.initials: M
authors.lastname: Laimer
authors.forename: A
authors.initials: A
authors.lastname: Klausegger
authors.forename: B
authors.initials: B
authors.lastname: Paulweber
authors.forename: B
authors.initials: B
authors.lastname: Kofler
authors.forename: H
authors.initials: H
authors.lastname: Hintner
authors.forename: J W
authors.initials: JW
authors.lastname: Bauer
mesh_terms: D000483:Alleles; D000963:Antimetabolites / Q000009:adverse effects; D001379:Azathioprine / Q000009:adverse effects; D005787:Gene Frequency; D006801:Humans; D008780:Methyltransferases / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012871:Skin Diseases / Q000188:drug therapy / Q000201:enzymology; D044465:White People / Q000235:genetics
chemical_list: D000963:Antimetabolites; D008780:Methyltransferases; C022745:thiopurine methyltransferase; D001379:Azathioprine