Human red cell ABO blood type antigen multi-PCR genotyping method and kit

A technology of ABO blood type and typing method, applied in the field of human red blood cell ABO blood group antigen multiple PCR typing method and kit, to achieve the effect of reducing the number of PCR reactions, high sensitivity, and improving detection throughput

Pending Publication Date: 2018-10-09
SHANGHAI BLOOD CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows individuals with an inherited or acquired type called AAO to test their own blood groups quickly without having to wait longer before getting tested because they don't have any other types that might be mistakenly added during this process. It also includes methods like multiplex polymerase chain reaction (PCR) and DNA sequencing techniques to make sure each individual does correctly identify different parts of these molecules. Overall, this new technique makes detecting ABoA typed red cells faster than traditional tests while reducing cost compared to current procedures.

Problems solved by technology

This patents describes how humans produce antihemophilial proteins called hemoglobin that help fight against harmful substances like thrombocytes. Hemolytic diseas include inherited bleeding due to defects in platelets caused by abnormalities during production processes. There may exist variations between these conditions leading to false alarms when diagnosing them based solely upon existing assay systems. However, multiplex polymerization techniques improve accuracy compared to traditional ones while reducing complexity.

Method used

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  • Human red cell ABO blood type antigen multi-PCR genotyping method and kit
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  • Human red cell ABO blood type antigen multi-PCR genotyping method and kit

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Embodiment 1

[0052] Embodiment 1 Human erythrocyte ABO blood group antigen multiplex PCR typing method of the present invention

[0053] (1) Method

[0054] Multiplex PCR based on PCR-SSP.

[0055] (2) Principle

[0056] The main inheritance mode of ABO blood type is: A, B, O are multiple alleles, in which A and B are dominant, and O is recessive. Design 4 pairs of specific primers to amplify O, non-O (ie A and B), B and non-B (ie A and O), respectively, and use beta-actin as an internal reference to form 2 multiplex PCR reactions. According to the presence or absence of the amplified band, the ABO blood type of the sample can be preliminarily judged (as shown in the table below).

[0057] Table 1 Comparison table of amplified bands and genotypes

[0058]

[0059] (3) Amplification primers

[0060]

[0061]

[0062] Note: The italics are the specific sites, and the bold underlines are the introduced mismatched bases.

[0063] Amplification primers are as follows: O and non-B...

Embodiment 2

[0069] Embodiment 2 Human erythrocyte ABO blood group antigen multiplex PCR typing kit of the present invention

[0070] A human erythrocyte ABO blood group antigen multiplex PCR typing kit, comprising the following primer pair;

[0071]

[0072] Instructions:

[0073] (1) Multiplex PCR is used for human erythrocyte ABO blood group antigen typing;

[0074] (2) The amplification system was as follows: O, non-B and internal reference primer pairs constituted a multiplex system; non-O, B and internal reference constituted a multiplex system; the amplification enzyme used Platinum Green Hot Start PCR Master Mix.

[0075] (3) The multiplex system of O, non-B and internal reference is as follows: DNA 2 microliters, non-B and internal reference primers 0.4 microliters each, O primer 0.15 microliters, Mix 10 microliters, ddH 2 O make up.

[0076] (3) The multiplex system of non-O, B and internal reference is as follows: 2 microliters of DNA, 0.08 microliters of non-O primers, 0....

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Abstract

The invention relates to a human red cell ABO blood type antigen multi-PCR genotyping method and a kit. The human red cell ABO blood type antigen multi-PCR genotyping method is characterized in that multi-PCR is adopted; four pair of specific primers are designed; O, no-O, B and no-B are correspondingly amplified; beta-actin is treated as an internal reference, and on that basis, two multi-PCR reactions systems are constructed; and the ABO blood types of a sample can be primarily determined on the basis of amplified bands. According to the method, the multi-PCR method is guided into ABO bloodtype genotyping; and the SNP loci of O allele and B allele are utilized to realize the ABO blood type genotyping. With the adoption of the method, the times of PCR reaction per person in ABO antigen genotyping can be reduced; the detection labor and the agent cost can be decreased; the detection throughput is increased; and the time needed in detection of a large quantity of samples can be shortened.

Description

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Claims

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Application Information

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Owner SHANGHAI BLOOD CENT
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