Multi-PCR-SBT genotyping method and reagent for ABO antigen of human erythrocyte blood type system

A PCR-SBT, genotyping method technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem that ABO genotyping cannot achieve high-throughput accurate typing and is difficult to adapt to Medical needs, etc.

Active Publication Date: 2019-04-02
浙江省血液中心
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Problems solved by technology

These defects directly lead to the inability of ABO genotyping to achieve high-throughput accurate typing, and it is difficult to adapt to the increasing medical needs in the future

Method used

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  • Multi-PCR-SBT genotyping method and reagent for ABO antigen of human erythrocyte blood type system
  • Multi-PCR-SBT genotyping method and reagent for ABO antigen of human erythrocyte blood type system
  • Multi-PCR-SBT genotyping method and reagent for ABO antigen of human erythrocyte blood type system

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Embodiment Construction

[0101] The content of the present invention will be described in further detail below in conjunction with the examples.

[0102] In this implementation, the content of the present invention will be described in detail by taking the blood of a blood donor as a test sample for typing the ABO antigen gene of the human blood group system as an example.

[0103] Grouping multiplex PCR-SBT method of the present invention comprises the following steps:

[0104] 1. Synthesize 14 amplification primers and 12 oligonucleotide sequencing primers. For the specific primer sequences, see the gene sequence in the content of the invention and the sequence list, which will not be repeated here. Dilute the amplification primers to 50 μM with pure water.

[0105] 2. Prepare 2 human genomic DNAs as templates for PCR amplification in subsequent steps.

[0106] 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 ki...

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Abstract

The invention belongs to the technical field of genotyping detection method, and in particular, relates to a multi-PCR-SBT genotyping method for an ABO antigen of a human erythrocyte blood type system. The invention also relates to a multi-PCR-SBT reagent for ABO antigen genotyping of the human erythrocyte blood type system. The reagent and method provided by the invention can be used as an independent and widely used identification method, solve the problem of rapid and accurate typing of an ABO antigen system, and exert the characteristics of high-throughput operation and accurate results ofmulti-PCR-SBT on ABO genotyping. Relevant applications in the fields of clinical blood transfusion medicine research and genetics can be highly valued, and important practical significance are achieved in research departments, and pharmaceutical research and reagent development departments.

Description

technical field [0001] The invention belongs to the technical field of genotyping detection methods, and in particular relates to a multiple PCR-SBT genotyping method for ABO antigens of human erythrocyte blood group system. The invention also relates to a multiplex PCR-SBT reagent for ABO antigen genotyping of human erythrocyte blood group system. Background technique [0002] The human ABO antigen is an important antigen in the ABO blood group system. The ABO gene is located on chromosome 9 (9q34.1-34.2), controlled by three multiple alleles A, B and O, and includes genes ranging in length from 28 to 688 bp. There are 7 exons and 6 introns with a length of about 19514bp, and the total length is about 18-20kb. The gene-encoded products are glycosyltransferases, which control the biosynthesis of ABO blood group antigens, thereby determining their blood type. [0003] The current most important method for ABO genotyping is PCR-SSP (PCR-sequence-specific primers), but this m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686C12N15/11
Inventor 朱发明应燕玲洪小珍何吉许先国陈舒马开荣
Owner 浙江省血液中心
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