A comparison of molecular and enzyme-based assays for the detection of thiopurine methyltransferase mutations

被引:33
作者
Coulthard, SA
Rabello, C
Robson, J
Howell, C
Minto, L
Middleton, PG
Gandhi, MK
Jackson, G
McLelland, J
O'Brien, H
Smith, S
Reid, MM
Pearson, ADJ
Hall, AG
机构
[1] Med Sch Newcastle Upon Tyne, Canc Res Unit, LRF Mol Pharmacol Specialist Programme, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England
[2] Royal Victoria Infirm, Dept Haematol, Newcastle Upon Tyne NE1 4LP, Tyne & Wear, England
[3] Royal Victoria Infirm, Dept Dermatol, Newcastle Upon Tyne NE1 4LP, Tyne & Wear, England
[4] Cumberland Infirm, Dept Haematol, Carlisle, England
[5] Torbay Hosp, Dept Haematol, Torquay, England
[6] Royal Victoria Infirm, Dept Child Hlth, Newcastle Upon Tyne NE1 4LP, Tyne & Wear, England
关键词
mutation analysis; leukaemia; mercaptopurine; thioguanine; pharmacogenetics;
D O I
10.1046/j.1365-2141.2000.02218.x
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
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.
引用
收藏
页码:599 / 604
页数:6
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