PCR-SBT (PCR-sequence-based typing) method and reagent for human blood platelet alloantigen system genotyping

Abstract

The invention provides a PCR-SBT (PCR-sequence-based typing) method for human blood platelet alloantigen system genotyping, comprising following steps: preparing a human genome DNA; amplifying a gene sequence of an antigen system in the human genome DNA; subjecting an amplification product to purification by double enzyme digestion; subjecting the purification product to sequencing PCR reaction; subjecting a sequencing product to purification by sodium acetate and ethanol precipitation, and carrying out capillary electrophoresis sequencing; subjecting an obtained sequence to software analysis to determine its genotype. The invention also provides a reagent used by the typing method. The method may serve as an independent and widely applicable identification method, the accurate typing problem of 28 antigen systems of HPA is solved, the feature that the PCR-SBT has high-throughput and accurate results for HPA genotyping operation is given to play, the method will be highly valued in related applications in the fields such as clinical blood transfusion medical research and genetics and has important practical significance to medical research institutes and pharmaceutical research and reagent development institutes.

Description

technical field [0001] The present invention relates to a genotyping detection method, in particular to a molecular biology detection method for antigen genotyping of a human platelet alloantigen system (Human Platelet Alloantigen, abbreviated as HPA) and reagents used in the method. Background technique [0002] There are complex blood group antigens on the surface of platelets. It not only has sugar chain antigens such as ABO, H, and Lewis shared with red blood cells, and HLA-I antigens shared with white blood cells, but also has a platelet-specific alloantigen system (humanplateletalloantigen, HPA), the HPA system shows a high degree of genetic polymorphism in the population. The differences in HPA antigens among individuals can produce antibodies through immune stimulation, and the antibodies can combine with the corresponding antigens of the donor to destroy platelets, resulting in ineffective platelet transfusion and other related diseases. Therefore, it is of great si...

AI Technical Summary

Problems solved by technology

However, the PCR-SBT typing method of HPA antigen gene at this stage is not perfect enough. Although some laboratories use the PCR-SBT method to genotype the HPA antigen, so far no systematic typing of the HPA1-28w system has been performed.

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