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Human HLA-B*5801 genotype kit and method

A technology of HLA-B and genotype, applied in the field of genetic diagnosis, can solve the problems of inability to perform stable and rapid batch operation, prone to pollution, and large operation differences, so as to avoid aerosol leakage, safety and pollution-free cost, specificity strong effect

Pending Publication Date: 2018-09-14
众福健康科技(杭州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is complicated to operate, takes a long time, and is prone to pollution. The operation of each batch is quite different, and it is impossible to perform stable and rapid batch operation.

Method used

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  • Human HLA-B*5801 genotype kit and method
  • Human HLA-B*5801 genotype kit and method
  • Human HLA-B*5801 genotype kit and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] The main components and accessories of the kit for rapid detection of human HLA-B*5801 genotype are as follows:

[0065]

[0066] The sequences of universal primers, universal fluorescent probes and connection probes are as follows:

[0067]

[0068]

[0069] The primer and probe sequences designed above are handed over to the synthesis company, synthesized by a DNA synthesizer, purified by HPLC, and a synthesis inspection report is issued. The sequence is the sequence provided by the present invention, but not limited to the sequence used in the present invention.

[0070] The above-mentioned universal fluorescent probe is a fluorescent double-labeled probe, which is labeled with FAM fluorescence at the 5' end and labeled with BHQ1-extracted fluorescein at the 3' end.

[0071] The dosage of the above-mentioned ligase is 1 U per person, the dosage of polymerase is 2 U per person, and the dosage of dNTP is 0.4 mmol / L.

[0072] This kit does not contain DNA ext...

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PUM

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Abstract

The invention provides a quick human HLA-B*5801 genotype detection kit and method. The kit mainly comprises universal primers, a universal fluorescent probe, ligation probes, ligation reaction and polymerization reaction mixed liquid, enzyme mixed liquid, a negative control and a positive control. Single nucleotide polymorphic site specificity is controlled through novel heatproof ligase reaction,only nucleotides matched with the ligation probes complete ligation reaction and start next polymerase chain reaction, and HLA-B*5801 genotype can be detected through the universal florescent probe;if the nucleotides are not matched with the ligation probes, ligation cannot be completed, and reaction is terminated. All detection processes can be completed in a same reaction system, and high specificity and high sensitivity in HLA-B*5801 genotype detection are achieved as compared with those of a traditional method.

Description

(1) Technical field [0001] The invention relates to a PCR kit for rapidly detecting human HLA-B*5801 gene and a method for detecting human HLA-B*5801 gene, belonging to the field of gene diagnosis. (2) Background technology [0002] Allopurinol is a hypoxanthine oxidase inhibitor, which can reduce the synthesis of uric acid and reduce the concentration of uric acid in the blood. It is the drug of choice for the treatment of gout. Clinically used for the treatment of: hyperuricemia; patients with chronic gout; tophi uric acid kidney stones or uric acid nephropathy. However, the drug is prone to cause severe skin adverse reactions, including mild drug eruption, severe exfoliative dermatitis, toxic epidermal necrolysis, etc., and has a high fatality rate. [0003] Human leukocyte antigen (HLA) is a group of genes located on chromosome 6. It is a gene complex with the highest allelic polymorphism among known genes so far. It is the most complex genetic polymorphism system in hu...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/156
Inventor 倪晓龙李亚波李春凤欧阳云
Owner 众福健康科技(杭州)有限公司
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