title: A comparison of molecular and enzyme-based assays for the detection of thiopurine methyltransferase mutations.
abstract: S-Methylation by thiopurine methyltransferase (TPMT) is an important route of metabolism for the thiopurine drugs. About one in 300 individuals are homozygous for a TPMT mutation associated with very low enzyme activity and severe myelosuppression if treated with standard doses of drug. To validate the use of molecular genetic techniques for the detection of TPMT deficiency, we have determined red blood cell TPMT activity in 240 adult blood donors and 55 normal children. Genotype was determined by restriction fragment length analysis of polymerase chain reaction products in a cohort of 79 of the blood donors and five cases of azathioprine-induced myelosupression, and this confirmed a close relationship between genotype and phenotype. In 17 of the 24 cases in which mutations were found, DNA was also available from remission bone marrow. In one of these cases, DNA from the remission marrow sample indicated the presence of a non-mutated allele that had not been seen in the blast DNA sample obtained at presentation. These results indicate that polymerase chain reaction-based assays give reliable and robust results for the detection of TPMT deficiency, but that caution should be exercised in relying exclusively on DNA obtained from lymphoblasts in childhood leukaemia.
authors.affiliation: The LRF Molecular Pharmacology Specialist Programme, Medical School, Newcastle Upon Tyne, UK.
authors.forename: S A
authors.initials: SA
authors.lastname: Coulthard
authors.forename: C
authors.initials: C
authors.lastname: Rabello
authors.forename: J
authors.initials: J
authors.lastname: Robson
authors.forename: C
authors.initials: C
authors.lastname: Howell
authors.forename: L
authors.initials: L
authors.lastname: Minto
authors.forename: P G
authors.initials: PG
authors.lastname: Middleton
authors.forename: M K
authors.initials: MK
authors.lastname: Gandhi
authors.forename: G
authors.initials: G
authors.lastname: Jackson
authors.forename: J
authors.initials: J
authors.lastname: McLelland
authors.forename: H
authors.initials: H
authors.lastname: O'Brien
authors.forename: S
authors.initials: S
authors.lastname: Smith
authors.forename: M M
authors.initials: MM
authors.lastname: Reid
authors.forename: A D
authors.initials: AD
authors.lastname: Pearson
authors.forename: A G
authors.initials: AG
authors.lastname: Hall
mesh_terms: D000293:Adolescent; D000328:Adult; D001379:Azathioprine / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use; D001782:Blood Donors; D002648:Child; D002675:Child, Preschool; D004252:DNA Mutational Analysis / Q000379:methods*; D005260:Female; D005838:Genotype; D006801:Humans; D007166:Immunosuppressive Agents / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use; D007223:Infant; D008297:Male; D015122:Mercaptopurine / Q000737:chemistry / Q000494:pharmacology; D008780:Methyltransferases / Q000097:blood / Q000172:deficiency* / Q000235:genetics*; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy / Q000201:enzymology / Q000235:genetics*; D012016:Reference Values; D012680:Sensitivity and Specificity; D013866:Thioguanine / Q000737:chemistry / Q000494:pharmacology
chemical_list: D007166:Immunosuppressive Agents; D015122:Mercaptopurine; D008780:Methyltransferases; C022745:thiopurine methyltransferase; D013866:Thioguanine; D001379:Azathioprine