Author/Editor		Ficko, T; Galvani, V; Rupreht, R; Dovč, T; Rožman, P
Title		Real-time PCR genotyping of human platelet alloantigens HPA-1, HPA-2, HPA-3 and HPA-5 is superior to the standard PCR-SSP method
Type		članek
Source		Transfus Med
Vol. and No.		Letnik 14
Publication year		2004
Volume		str. 425-32
Language		eng
Abstract		Genotyping of the human platelet alloantigens (HPA) is useful for the diagnosis and therapy of the patients with alloimmune thrombocytopenic syndromes, such as post-transfusion refractoriness to platelets, post-transfusion thrombocytopenic purpura and foetomaternal alloimmune thrombocytopenia. We have developed, optimized and validated a new method for simultaneous genotyping of HPAs - HPA-1, HPA-2, HPA-3 and HPA-5 - by using the real-time polymerase chain reaction (PCR) based on TaqMan technology. Its performances were compared to those of the standard PCR-sequence-specific primers (SSP) method by testing 120 DNA samples. Several discrepancies between the two methods have been observed, especially in the HPA-3 genotyping. Evidently, the PCRSSP method produced several false positive results due to its technical drawbacks. Based on our comparison, we believe that the new real-time TaqMan PCR assay for the HPA-1, HPA-2, HPA-3 and HPA-5 genotyping is faster, more reliable and reproducible, compared to the standard PCR-SSP.
Descriptors		ANTIGENS, HUMAN PLATELET THROMBOCYTOPENIA GENOTYPE BLOOD DONORS POLYMERASE CHAIN REACTION SEQUENCE ANALYSIS, DNA