Genotyping of human platelet antigen-15 by single closed-tube Tm-shift method.
| Autor: | Zhou SH; Dalian Blood Center, Dalian, China., Liu M, An WX, Liang XH, Yu WJ, Gong BL, Piao FY |
|---|---|
| Jazyk: | angličtina |
| Zdroj: | International journal of laboratory hematology [Int J Lab Hematol] 2012 Feb; Vol. 34 (1), pp. 41-6. Date of Electronic Publication: 2011 Jun 13. |
| DOI: | 10.1111/j.1751-553X.2011.01344.x |
| Abstrakt: | Introduction: Genotyping of human platelet antigens (HPA) is useful for the diagnosis and prevention of platelet alloimmune syndromes. HPA-15 might play an important role in the development of platelet alloimmune syndromes. There are several disadvantages in the conventional methods for HPA-15 genotyping. The aim of this study was to develop a new method for HPA-15 genotyping by using single closed-tube melting temperature (T(m))-shift genotyping. Methods: Two GC-rich tails of different lengths were attached to 5'-end of HPA-15 allele-specific PCR primers, such that HPA-15 alleles can be discriminated by the T(m)s of the PCR products. One hundred blood samples were genotyped for HPA-15 by the T(m)-shift and conventional polymerase chain reaction with sequence-specific primers (PCR-SSP). Results: The comparison of the PCR-SSP and the T(m)-shift method showed four discordant results in one hundred samples tested. Confirmatory results demonstrated that the PCR-SSP produced several errors, whereas HPA-15 genotyping by T(m)-shift is correct. The retesting results of T(m)-shift method were consistent with those of the initial testing. Conclusion: The single closed-tube T(m)-shift method for HPA-15 genotyping is high-throughput, rapid, reliable, reproducible and cost-effective and it is superior to conventional PCR-SSP used in routine genotyping of HPA-15. (© 2011 Blackwell Publishing Ltd.) |
| Databáze: | MEDLINE |
| Externí odkaz: |
|