Detection of thiopurine s-methyltransferase mutations by template directed determinator in incorporation with fluorescence polarization (TDI-FP)

摘要

Objective: To develop a new method for the detection of TPMT gene mutations and determine the frequencies of four TPMT alleles, TPMT ~*1, ~*3A , ~*3B and ~*3C in a healthy Chinese population. Methods: A TDI-FP assay system was set up in out lab. To evaluate this system, 220 healthy individuals were analyzed for the polymorphic sites at positions 460(G→A)and 719(A→G)of the TPMP gene using our new TDI FP method. Results: Three TPMP * 3C(G~(460)→G~(719)) heterozygotes were identified, TPMP ~*3A and TPMP ~*3B were not found. All mutations were confirmed by conventional DNA sequencing analysis. Conclusion: TDI-FP method has proven to be very efficient as a rapid and accurate approach for TPMP genotyping. TPMP ~*3C was the only polymorphism identified in this clinical samples we have registered.