A pcr-sbt method and reagent for genotyping of human platelet alloantigen system

An alloantigen system, PCR-SBT technology, applied in the field of genotyping detection, can solve the problems of incomplete typing methods, unsystematic typing of HPA1-28w system, etc.

Active Publication Date: 2017-12-19
浙江省血液中心
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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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  • A pcr-sbt method and reagent for genotyping of human platelet alloantigen system
  • A pcr-sbt method and reagent for genotyping of human platelet alloantigen system
  • A pcr-sbt method and reagent for genotyping of human platelet alloantigen system

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Embodiment Construction

[0173] The content of the present invention will be described in further detail below in conjunction with the examples.

[0174] In this implementation, the genotyping of the HPA1-28w system antigen gene is performed using the blood of a blood donor as a test sample as an example to describe the content of the present invention in detail.

[0175] The PCR-SBT method of genotyping of the present invention comprises the following steps:

[0176] 1. Prepare human genomic DNA as a template for PCR amplification in subsequent steps.

[0177] 200 µl of whole blood to be tested was taken, and genomic DNA was extracted according to the instructions of the QuickGene DNA whole blood kit S kit, and the concentration and purity of the genome were determined by a spectrophotometer.

[0178] 2. Synthesize 18 pairs of amplification primers. For the specific primer sequences, please refer to the gene sequence in the content of the invention and the sequence list, which will not be repeated h...

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Abstract

The invention provides a PCR-SBT method for genotyping human platelet alloantigen system, the method comprising the following steps: preparing human genome DNA; amplifying the gene sequence of the antigen system in human genome DNA; performing double-enzyme synthesis on the amplified product The purified product was subjected to sequencing PCR reaction; the sequencing product was purified by sodium acetate-ethanol precipitation method, followed by capillary electrophoresis sequencing; the obtained sequence was analyzed by software to determine its genotype. The present invention also provides reagents used in the above typing method. The present invention can be used as an independent and widely used identification method, solves the problem of accurate typing of 28 antigen systems of HPA, utilizes the characteristics of high-throughput and accurate results of PCR-SBT for HPA gene typing operation, and can be used in clinical blood transfusion medicine research The related applications in fields such as genetics and genetics will be highly valued, and it has important practical significance for medical research units, pharmaceutical research and reagent development units.

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 (HPA) system and reagents used in the method. Background technique [0002] There are complex blood group antigens on the surface of platelets, which not only have 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 have a platelet-specific alloantigen system (human platelet alloantigen, 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 significance to study the platelet allo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 朱发明何吉陈舒洪小珍许先国刘瑛吕杭军
Owner 浙江省血液中心
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