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 syste...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.展开更多
Objective: To develop a new high sensitivity, rapid and simple mycobacterium tuberculosis DNA detection method using fluorescence polarization technology. Methods: In our asymmetric PCR protocol, 100 times mole of TB-...Objective: To develop a new high sensitivity, rapid and simple mycobacterium tuberculosis DNA detection method using fluorescence polarization technology. Methods: In our asymmetric PCR protocol, 100 times mole of TB-R primer was added than in the usual symmetric PCR to get enough single strands PCR product. The probe TB-5′-TAMRA and PCR products were mixed in a tube and incubate for 5-15 min at 46 ℃.The polarization (mp) was measured using victor2 Multilabel Plate Reader. Results: Asymmetric and symmetric PCR products were analyzed by the FP method. Asymmetric PCR products are detected more sensitively than symmetric ones. The polarization values of probe associated with asymmetric products were much higher than with symmetric products. Conclusion: This fluorescence polarization assay in conjunction with asymmetric PCR is a powerful and widely applicable method for the rapid and sensitive detection of micro-organisms in clinical laboratories.展开更多
文摘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.
文摘Objective: To develop a new high sensitivity, rapid and simple mycobacterium tuberculosis DNA detection method using fluorescence polarization technology. Methods: In our asymmetric PCR protocol, 100 times mole of TB-R primer was added than in the usual symmetric PCR to get enough single strands PCR product. The probe TB-5′-TAMRA and PCR products were mixed in a tube and incubate for 5-15 min at 46 ℃.The polarization (mp) was measured using victor2 Multilabel Plate Reader. Results: Asymmetric and symmetric PCR products were analyzed by the FP method. Asymmetric PCR products are detected more sensitively than symmetric ones. The polarization values of probe associated with asymmetric products were much higher than with symmetric products. Conclusion: This fluorescence polarization assay in conjunction with asymmetric PCR is a powerful and widely applicable method for the rapid and sensitive detection of micro-organisms in clinical laboratories.
