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Human RhD blood group genotyping detection primer set and application

A technology for genotyping and detecting primers, which is applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc.

Inactive Publication Date: 2019-04-19
江苏中济万泰生物医药有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

The common types in Chinese include Weak RhD 15 and DEL RhD1227A, as well as the allele detection of RhD exons 1-7, 9-10 and the four common Rh blood group antigen gene polymorphisms of RhC, Rhc, RhE, and Rhe. However, the prior art lacks a comprehensive and holistic assay kit for human RhD blood group genotyping detection

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  • Human RhD blood group genotyping detection primer set and application
  • Human RhD blood group genotyping detection primer set and application
  • Human RhD blood group genotyping detection primer set and application

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Embodiment

[0025] A kit for detecting human RhD blood type genotyping, comprising primers of SEQ ID No. 1-30, dNTP-Buffer, cresyl red sodium salt and SYBR GreenⅠ. The detection of each person includes 15 detection wells, in addition to placing a pair of allele primers in each well, it also includes internal reference quality control primers, the sequences of which are shown in SEQ ID No.31 and SEQ ID No.32.

[0026] Storage conditions and expiration date

[0027] All components of the kit should be stored below -18°C, and the validity period is 6 months.

[0028] Sample request:

[0029] 1. The sample required for this kit is whole blood, which needs to be extracted from whole blood DNA, and its DNA sample concentration needs to be determined.

[0030] 2. When extracting DNA from whole blood, it is not allowed to use heparin anticoagulated whole blood samples, and EDTA anticoagulant is recommended.

[0031] 3. The DNA sample concentration should be 40-70ng / μl, and the purity A260 / A280...

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Abstract

The invention discloses a human RhD blood group genotyping detection primer set which comprises 16 pairs of primers shown in SEQ ID No. 1-32. Based on the polymerase chain reaction (fluorescence PCR)principle, the first base at the 3' terminals of the primers is complementary to allele-specific base, and the specific primer pairs only amplify alleles matched with them. In primer design, accordingto human gene sequences disclosed by a blood group antigen gene mutation database (dbRBC) in a DNA sequence database (Genbank) established by the US National Center for Biotechnology Information (NCBI), suitable specific mutational sites are selected to design the primers; in a PCR dye incorporation method, a fluorescent dye is incorporated into an amplification product of DNA to show the DNA amplification effect; melting curve analysis is conducted after amplification; whether the target product is specifically amplified or not can be determined through melting curve analysis, and accordingly alleles are judged.

Description

technical field [0001] The invention relates to the technical field of gene diagnosis products, in particular to a human RhD blood type genotyping detection primer set and its application. Background technique [0002] The Rh blood group is the most complex and polymorphic system in the human red blood cell blood group system, and it is also the main red blood cell blood group that causes clinical transfusion reactions and severe hemolytic disease of the newborn. At present, more than 50 kinds of Rh blood group antigens have been found, among which the RhD antigen has strong immunogenicity and is encoded by the RhD gene, which is the focus of blood group research. The common types in Chinese include Weak RhD 15 and DEL RhD1227A, as well as the allele detection of RhD exons 1-7, 9-10 and the four common Rh blood group antigen gene polymorphisms of RhC, Rhc, RhE, and Rhe. However, the prior art lacks a comprehensive and integral assay kit for human RhD blood group genotyping ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 林裕翔张浩钧朱晓洁
Owner 江苏中济万泰生物医药有限公司
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