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Multiple PCR detection method and kit for human platelet blood type

A technology of human platelets and kits, applied in biochemical equipment and methods, microbial measurement/inspection, recombinant DNA technology, etc., can solve the problems of low cost and large throughput, and achieve low cost, large throughput, and interpretation clear effect

Active Publication Date: 2019-12-06
SHANGHAI BLOOD CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are no reports on HPA genotyping methods and related kits with accurate and reliable typing results, high sensitivity, good specificity, high throughput, low cost, and high efficiency.

Method used

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  • Multiple PCR detection method and kit for human platelet blood type
  • Multiple PCR detection method and kit for human platelet blood type
  • Multiple PCR detection method and kit for human platelet blood type

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Use the kit and method of the present invention to carry out HPA genotyping, and verify the typing accuracy, specificity and sensitivity of the kit of the present invention.

[0038] 1. Primers

[0039] The primers used in the present invention were all synthesized by Shanghai Sangong Company and purified by UltraPage purification. Primer sequences are shown in Table 1.

[0040] Table 1 Primer Sequence

[0041]

[0042]

[0043] 2. Blood DNA Extraction

[0044] Take 1mL of whole blood (EDTA anticoagulant), use blood genomic DNA extraction kit (Beijing Tiangen Company, DP318) to extract genomic DNA according to the manufacturer's instructions, and use Nanodrop 2000 (Thermo) to determine A 260 OD value and A 260 :A 280 ratio.

[0045] 3. SSP-PCR genotyping of samples

[0046] Prepare 2×primer master mix according to Table 2, add 5 μL of 2×primer master mix, 5 μL of 2×Taq enzyme reaction master mix (including about 0.5U of Taq enzyme, PCR reaction buffer and a...

Embodiment 2

[0065] A human platelet blood group antigen genotyping kit, comprising the following reagents: primers of SEQ ID NO: 1-23.

Embodiment 3

[0067] A human platelet blood group antigen genotyping kit, comprising the following reagents: primers of SEQ ID NO: 1-23. Specifically, the primer premix is ​​included, and the concentration of each primer in each tube of the primer premix is ​​shown in the table below:

[0068]

[0069]

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PUM

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Abstract

The invention provides a multi-PCR (polymerase chain reaction) detection method and a reagent kit for blood types of human blood platelets. The reagent kit comprises primers with sequences shown as SEQ ID NO:1-23. Appropriate PCR systems and reaction conditions are recorded, and the reagent kit can be used for carrying out genetic typing on antigens HPA1-6 and HPA15 of the blood types of the blood platelets. The multi-PCR detection method and the reagent kit for the blood types of the human blood platelets have the advantages of reliable typing results, high sensitivity, throughput and efficiency, good specificity, capability of carrying out multi-PCR simplicity, convenience and speediness in operation, low cost and broad application prospects in the aspects of diagnosis of alloimmune thrombopenia, establishment of compatible blood platelet donor banks and extensive survey on population HPA (heparanase) gene frequencies.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a multiplex PCR detection method and kit for human platelet blood type. Background technique [0002] A variety of platelet antigens (HPA) are distributed on the surface of human platelets, which constitute the blood group system of platelets. Thirty-five platelet blood group antigens have been identified serologically to date, and due to SNP variations between these HPA alleles, resulting in the formation of distinct epitopes that can be recognized by alloimmunization, platelet alloimmunization produces the corresponding platelet Antibody. Among them, the corresponding antibodies against HPA1-6 and HPA15 antigens are the main cause of clinically relevant platelet alloimmunization, which can cause platelet blood group antigen immune-related clinical conditions, such as neonatal alloimmune thrombocytopenia (NAIT), Post-transfusion purpura (PTP) and platelet transfusion ineffective. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6881C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6881C12Q2600/156C12Q2600/16C12Q2537/143
Inventor 朱慧君陆萍傅敏李睿书
Owner SHANGHAI BLOOD CENT
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