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.
Transfus Med
; 14(6): 425-32, 2004 Dec.
Article
em En
| MEDLINE
| ID: mdl-15569237
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 PCR-SSP 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.
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Base de dados:
MEDLINE
Assunto principal:
Reação em Cadeia da Polimerase
/
Antígenos de Plaquetas Humanas
/
Polimorfismo Conformacional de Fita Simples
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Revista:
Transfus Med
Assunto da revista:
HEMATOLOGIA
Ano de publicação:
2004
Tipo de documento:
Article
País de afiliação:
Eslovênia