Para-bombay phenotype FUT1 508dupT allele as well as detection method and application thereof

A technology of alleles and detection methods, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems that ordinary laboratories cannot carry out, expensive reagents, high technical requirements, etc., to solve the problem of difficult blood transfusion, low screening cost, The effect of low technical requirements

Pending Publication Date: 2020-10-23
沈阳中心血站
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

At present, the identification of FUT1 gene mutations is carried out by sequencing methods, but this method has some insurmountable shortcomings: complex operation process, high technical requirements, expensive reagents, high detection costs, low penetration rate of sequencers, and cannot be carried out by ordinary laboratories etc. These shortcomings make it very limited in clinical application

Method used

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  • Para-bombay phenotype FUT1 508dupT allele as well as detection method and application thereof
  • Para-bombay phenotype FUT1 508dupT allele as well as detection method and application thereof
  • Para-bombay phenotype FUT1 508dupT allele as well as detection method and application thereof

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Embodiment 1

[0038] Example 1 Bombay blood type FUT1 508dupT allele detection method.

[0039] 1. Preparation of DNA template.

[0040] Whole blood genomic DNA was extracted using purchased kits, and the specific steps were as follows.

[0041] (1) Take 200 μL EDTA anticoagulated whole blood sample, add 20 μL Proteinase K solution, and mix well.

[0042] (2) Add 200 μL buffer GB, mix thoroughly by inversion, and place at 56°C for 10 minutes, during which, invert and mix several times, the solution should become clear and bright.

[0043] (3) Add 200 μL of absolute ethanol, mix thoroughly by inversion, flocculent precipitation may appear at this moment.

[0044] (4) Transfer the solution and the flocculent precipitate obtained in the previous step into an adsorption column CB3. Centrifuge at 12000rpm for 30sec, pour off the waste liquid in the collection tube, and put the adsorption column CB3 into the collection tube.

[0045] (5) Add 500 μL of buffer GD to the adsorption column CB3, c...

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Abstract

The invention belongs to the technical field of molecular biological analysis and detection, and relates to a para-bombay phenotype FUT1 508dupT allele as well as a detection method and application thereof. The para-bombay phenotype FUT1 508dupT allele is formed by repeating a T base from the 508th site of an initiation codon in an Hh blood type system variant FUT1 gene coding region. The invention also discloses an application of the para-bombay phenotype FUT1 508dupT allele in the preparation of red blood cell para-bombay phenotype products. A method for detecting the para-bombay phenotype FUT1 508dupT allele comprises the following steps: performing PCR amplification on DNA of a blood sample to be detected by using primers to obtain a specific amplification band pattern, comparing the specific amplification band pattern with an FUT1 wild type amplification band pattern, and determining whether the sample to be detected has the para-bombay phenotype FUT1 508dupT allele or not. The detection method not only can be used for identifying the FUT1 gene of specific individuals and identifying bombay (para-bombay) phenotype, but also can be used for screening a large number of samples due to low screening cost and simple operation so as to establish a bombay (para-bombay) phenotype donator database and provide matched blood for bombay (para-bombay) phenotype patients.

Description

technical field [0001] The invention belongs to the technical field of molecular biology analysis and detection, and relates to a Mumbai-like FUT1 508dupT allele and its detection method and application, in particular to a detection method of a known mutant gene doped in a wild-type gene cluster. Background technique [0002] The blood group system refers to the classification of blood according to the differences in the antigens on the red blood cell membrane. There are 39 human erythrocyte blood group systems discovered so far. The Hh blood group system, also known as the Bombay blood group system, is a human blood group system that classifies blood according to the presence or absence of H antigen on the surface of red blood cells. The H antigen is the precursor of each ABO blood group antigen. The FUT1 gene, which determines the H antigen, is located on chromosome 19 and is more than 5000 base pairs in length and contains 4 exons. The FUT1 gene has two alleles H and h...

Claims

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Application Information

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IPC IPC(8): C12Q1/6876C12Q1/6858C12N15/12
CPCC12Q1/6876C12Q1/6858C12Q2600/156C12Q2600/166C12Q2531/113C12Q2545/101C12Q2565/125
Inventor 李剑平林凤秋李晓丰王宏阳
Owner 沈阳中心血站
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