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Detection kit for hla-b5801 allele and its application

A technology of HLA-B5801, 1. HLA-B5801, applied in the field of detection kits for in situ genes, can solve the problems of complex qPCR reaction system and unstable results, and achieve the benefits of popularization and popularization, simple use and high accuracy Effect

Active Publication Date: 2021-12-07
广东辉锦创兴生物医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the richness of the HLA-B5801 allele sequence, the existing qPCR method can only use multiple primers to cover the entire sequence of the HLA-B5801 allele, resulting in a complex qPCR reaction system and unstable results

Method used

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  • Detection kit for hla-b5801 allele and its application
  • Detection kit for hla-b5801 allele and its application
  • Detection kit for hla-b5801 allele and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1. Sequence Alignment Analysis of Existing HLA-B5801 Genotypes

[0017] In order to overcome the above-mentioned defects in the qPCR method, the present invention provides a pair of primers with degenerate bases, which can cover the HLA-B5801 genotype sequence to the greatest extent while ensuring the minimum degeneracy. And through the combination of the TaqMan probe and the degenerate primer, the HLA-B5801 allele can be accurately detected in one qPCR reaction.

[0018] Currently known B-58:01 has the following subtypes: B*58:01:01:01, B*58:01:01:02, B*58:01:02, B*58:01:03, B *58:01:04, B*58:01:05, B*58:01:06, B*58:01:07, B*58:01:08, B*58:01:09, B*58 :01:10, B*58:01:11, B*58:01:12, B*58:01:13, B*58:01:14, B*58:01:15, B*58:01 :16, B*58:01:17, B*58:01:18, B*58:01:19, and B*58:01:20.

[0019] In order to make the designed fluorescent quantitative PCR cover all the above allelic loci, the inventors compared and analyzed the sequences of the above subtypes, and ...

Embodiment 2

[0028] Example 2. Sample HLA-B5801 Positive / Negative Detection

[0029] For clinical samples of peripheral whole blood of the sample type detected in the present invention, after the genomic DNA is extracted from the blood sample, the sequences of different subtypes of HLA-B5801 alleles are obtained by Sanger sequencing.

[0030] The reaction was carried out through the following fluorescent quantitative PCR reaction system. The qPCR reaction system used the product of Beijing Kangwei Century Biotechnology Company, as shown in Table 3, and the reaction conditions were shown in Table 4.

[0031] Table 3. Reaction system for detecting HLA-B5801 alleles by qPCR

[0032]

[0033] Table 4. qPCR reaction conditions

[0034]

[0035]

[0036] On the fluorescent quantitative PCR instrument ABI7500, the fluorescent quantitative reaction curves of HLA-B5801 positive and negative detection can be found in figure 1 .

[0037] 20 cases of HLA-B5801-positive and 20 cases of HLA-...

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Abstract

The present invention relates to the detection kit of HLA-B5801 allele and application thereof, and the kit of the present invention comprises the qPCR primer and probe of detecting HLA-B5801 allele, uses primer and probe of the present invention to overcome existing qPCR method Only multiple primers and probes can be used to cover the entire sequence of the HLA-B5801 allele, resulting in a complex qPCR reaction system and unstable results. The kit of the invention has the advantages of simple use, high accuracy, monitorable process, rapid result, and no electrophoresis process, which is beneficial to clinical promotion and popularization.

Description

technical field [0001] The invention relates to the field of molecular diagnosis, in particular to a detection kit for HLA-B5801 allele and application thereof. Background technique [0002] Hyperuricemia is a metabolic disease caused by abnormal purine metabolism, excessive production of blood uric acid or excretion disorder. It is not only the main risk factor of gout, but also an important part of metabolic syndrome, which affects the functions of the heart, kidneys and other organs. It is an independent risk factor for hypertension and coronary heart disease, and an important risk factor for chronic renal insufficiency, causing serious harm to human health. [0003] Xanthine oxidase inhibitors such as allopurinol can reduce the synthesis of uric acid in the body, and are clinically used in the prevention and treatment of diseases such as hyperuricemia, gout and gouty arthritis. Allopurinol is the most suitable drug for primary and secondary gout patients with excessive ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6851
CPCC12Q1/6851C12Q2563/107C12Q2545/101C12Q2545/113
Inventor 胡锦张凤笑
Owner 广东辉锦创兴生物医学科技有限公司
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