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Primer combination for detecting Diego blood group genotyping of human red cells and kit

A red blood cell and blood type technology, which is applied in the field of kits for detecting human red blood cell Diego blood type genotyping, can solve the problems of difficult purchase and high price of reagents

Inactive Publication Date: 2015-06-10
天津市秀鹏生物技术开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is: to overcome the high price of serological detection reagents, the deficiencies such as reagents are not easy to buy, provide a kind of immunology method that can simply and conveniently detect the primer set of human erythrocyte Diego blood group gene

Method used

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  • Primer combination for detecting Diego blood group genotyping of human red cells and kit
  • Primer combination for detecting Diego blood group genotyping of human red cells and kit
  • Primer combination for detecting Diego blood group genotyping of human red cells and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] According to the Diego allele sequence released by the National Center for Biotechnology Information (NCBI Gene Bank), a primer set for detecting the Diego blood group gene of human erythrocytes was designed, which includes Di a Primer pair, Di b Primer pairs and internal reference primer pairs.

Embodiment 2

[0021] A test kit for detecting the Diego blood group gene of human erythrocytes, comprising a Di a Primer pair, Di b Primer plates and enriched dNTP-Buffer for primer pairs and internal reference primer pairs.

[0022] The preparation method of the test kit for detecting human erythrocyte Diego blood group gene is:

[0023] (1) According to the specific primer hole map of human erythrocyte Diego blood type gene (see figure 1 ), respectively Di a Primer pair, Di b Primer pairs and internal reference primer pairs are coated on the corresponding positions of the primer plate and dried;

[0024] (2) Prepare 440μl concentrated dNTP-Buffer according to the mixture of 220mM dNTP, 3.5mM Mg2+, 500mM KCL, 100mM Tris-HCL, 1% TritonX-100;

[0025] (3) The primer plate coated with the primer pair includes 2 specific primer wells per person, and the dNTP-Buffer is concentrated to form a kit for detecting the Diego blood group gene of human erythrocytes.

Embodiment 3

[0027] Use operational procedures.

[0028] 1. Preparation of 2.5% agarose gel.

[0029] 1. Add 2.5g agarose to 100ml 1×TBE solution (Tris-borate-EDTA solution) and heat until a uniform gel solution is formed. Then add an appropriate amount of electrophoresis dye and mix well.

[0030] 2. Place the gel tank in a horizontal position, add an appropriate amount of gel solution into the gel tank, and insert the electrophoresis comb. Move the gel tank horizontally back and forth to ensure even coverage of the gel solution.

[0031] 3. After the gel is completely solidified, pull out the electrophoresis comb vertically.

[0032] 2. The PCR cycle parameters are shown in Table 1.

[0033] Table 1 Characteristic table of amplification parameters.

[0034] 1 96℃ / 2min 1 cycle 2 96℃ / 20sec, 68℃ / 60sec 5 cycles 3 96°C / 20 sec, 65°C / 45 sec, 72°C / 30 sec 10 cycles 4 96°C / 20 sec, 62°C / 45 sec, 72°C / 30 sec 15 cycles 5 72℃ / 3min 1 cycle 6 Stor...

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Abstract

The invention discloses a primer combination for detecting the Diego blood group genotyping of human red cells and a kit. The primer combination for detecting Diego blood group genotyping of human red cells comprises a Dia primer pair, a Dib primer pair and an internal primer pair. According to the kit provided by the invention, various limitations on Diego blood type determined by a serologic method can be overcome, therefore the primer combination is an indispensable assistant means of accurately judging the blood type, and has an extensive application prospect and a clinical reference value.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a kit (PCR-SSP method) for detecting the genotyping of human erythrocyte Diego blood group. Background technique [0002] The corresponding ISBT symbols of traditional Dia and Dib are DI1 and DI2, which are the most clinically significant pair of antigens in the Diego blood group system. Anti-Dia corresponding to the antigens of the Diego blood group system can cause hemolytic disease of the newborn and can also destroy the imported Dia antigen erythrocytes, anti-Dib are very rare, but they can also cause hemolytic disease of the newborn (HDN) and hemolytic transfusion reactions, which have clinical significance. Dia and Dib antigens can be identified by serological methods, but the reagents are relatively expensive, especially anti-Dib. -Dib typing reagents are even more difficult to purchase in the market, and it is very difficult to find a suitable blood source for clinical blood ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6876C12Q2600/166
Inventor 赵卫军
Owner 天津市秀鹏生物技术开发有限公司
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