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Kit used for detecting human platelet specific antigen HPA 1-29 gene typing

A genotyping and kit technology, applied in the field of biomedical clinical molecular detection, can solve the problems of not being suitable for wide application and high cost, and achieve the effect of being suitable for large-scale detection, high-efficiency detection requirements, and improving accuracy and sensitivity

Active Publication Date: 2020-06-05
JIANGSU WEIHE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It uses MGB probes and FAM / VIC to mark the a / b genotypes of HPA subtypes respectively, and judges the allele type according to the two fluorescence curves after amplification. This method is costly and requires a / b for each subtype Two MGB probes, not suitable for wide application

Method used

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  • Kit used for detecting human platelet specific antigen HPA 1-29 gene typing
  • Kit used for detecting human platelet specific antigen HPA 1-29 gene typing
  • Kit used for detecting human platelet specific antigen HPA 1-29 gene typing

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Experimental program
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Effect test

Embodiment 1

[0041] 1. Raw materials and equipment

[0042] 1.1 Preparation of PCR reaction mixture: 0.18mM deoxynucleotide (dNTP), 1.8mM magnesium chloride (MgCl2), 60.3mM potassium chloride (KCl), 18.9mM trishydrochloride (Tris-HCl) , glycerol (glycerol) 0.6% (v / v), dimethyl sulfoxide (DMSO) 5% (v / v) and formamide 2.5% (v / v), prepare 6*PCR reaction mixture 50mL according to the above ratio , Long-term storage at -20°C, temporary storage at 4°C for later use.

[0043] 1.2 Preparation of reaction plate:

[0044] 1.2.1 Preparation of primer-probe mixture: primer 0.54OD / mL, probe 1.05μM, make 6*primer-probe mixture.

[0045] 1.2.2 Click the board according to the table:

[0046]

[0047]

[0048]

[0049] The nucleotide sequence of SEQ ID No.001~110 is shown in the following table:

[0050]

[0051]

[0052]

[0053]

[0054] 1.2.3 Freeze-drying of the reaction plate: freeze-dry the reaction plate according to the procedure in the table below. Packed in a light-proo...

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Abstract

The invention provides a PCR (Polymerase Chain Reaction) amplification primer group and Taqman probe combination used for detecting HPA (Human Platelet Antigen) 1-29 gene typing. The combination comprises 16 primer groups. The invention also provides a kit containing the PCR amplification primer group and Taqman probe composition. The invention has the following technical effects that an ARMS (Amplification refractory mutation system) analysis method is adopted and is combined with a Taqman multiplex fluorescence PCR technology to carry out qualitative typing detection on HPA 1-29 genes, an improvement is made on the basis of a traditional ARMS, various defects of an existing detection method are overcome, and therefore, the invention has a wide application prospect and a high clinical reference value.

Description

technical field [0001] The invention relates to a method and a kit for detecting human platelet-specific antigen HPA 1-29 genotyping, belonging to the field of biomedical clinical molecular detection. Background technique [0002] Platelet is one of the formed components in mammalian blood, with irregular shape and no nucleus. Its main function is blood coagulation and hemostasis, and plays an important role in inflammatory response. There are a large number of antigens on the surface of platelets, which are usually divided into two categories: one is antigens shared with other blood cells or tissues, called platelet-associated antigens, such as HLA-I antigens, ABO antigen system, etc.; the other is platelet-specific antigens , showing the unique genetic polymorphism of platelets, that is, human platelet antigen (Human Platelet Antigen, HPA). [0003] HPA is an antigenic epitope located on the platelet membrane glycoprotein (glycoprotein, GP), and is an inherent antigen of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 陈炤源王浩张金涛章婷婷顾逸飞
Owner JIANGSU WEIHE BIOTECH