Detection method and kit of HLA (Human Leukocyte Antigen)-B*58:01 allele

An HLA-B, allele technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of cumbersome operation, high cost, and long time.

Inactive Publication Date: 2014-05-21
GUANGDONG UNITY BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, while having many advantages, this method also has disadvantages such as cumbersome operation, long time consumption and high cost.
In particular, the shortcomings of cumbersom...

Method used

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  • Detection method and kit of HLA (Human Leukocyte Antigen)-B*58:01 allele
  • Detection method and kit of HLA (Human Leukocyte Antigen)-B*58:01 allele
  • Detection method and kit of HLA (Human Leukocyte Antigen)-B*58:01 allele

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] HLA-B*58:01 allele detection kit (SSP-PCR fluorescent probe method) and its use.

[0058] (1) Prepare a kit including the following components: 1 tube of reaction solution 1 (650μl / tube), 1 tube of reaction solution 2 (650μl / tube), 1 tube of reaction solution 3 (650μl / tube), EZ Taq enzyme mixture (150μl / tube) 1 tube, standard solution 1 (30μl / tube), standard solution 2 (30μl / tube).

[0059] (2) Sample collection, transportation and storage: use a disposable vacuum blood collection device, perform aseptic operation, take blood samples from individuals to be tested, and place the blood samples in a low-temperature refrigerator (-70°C or -20°C) after labeling Store in the freezer. For centralized inspection, specimens must be transported in an environment below 0°C and delivered to the laboratory within 24 hours.

[0060] (3) Inspection steps and result analysis:

[0061] Take 300 μl of the blood sample to be tested, extract the DNA in the blood sample (deleted) and standard 1 a...

Embodiment 2

[0071] Example 2: Results and analysis of 30 clinical blood samples detected by the kit of the present invention

[0072] Using the single-blind experiment method, 30 cases were selected from 500 blood samples from Chongqing Blood Center to test with the kit of the present invention, using TBG’s HLAssure ABCDRDQ SBT sequencing kit, using PCR-SBT gold standard method kit Review the results of the experiment. The test data showed that the PCR amplifications of the 30 clinical blood samples tested by this kit were all effective amplifications. Except for No. 1 sample amplification system I and No. 6 sample amplification system III, the target sequences of control genes in other samples were all amplified; except for the amplification system I (Ct34.13) of sample No. 10 and sample No. 17 Except for amplification system I (Ct34.93) and III (Ct34.54), the CT values ​​of the target sequences of the control genes in other samples were all less than 30. Among them, 8 samples had HLA-B*5...

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Abstract

The invention relates to a detection method of HLA (Human Leukocyte Antigen)-B*58:01 allele. The detection method comprises the following steps: providing a mixture of a sample to be tested, a nucleic acid amplification system and a fluorescence detection system; circularly amplifying target polynucleotide through an amplified reaction; indirectly combining a fluorescence generating group and an amplified target polynucleotide sequence; and detecting the fluorescent amount generated by the fluorescence generating group so as to determine existence of the target polynucleotide and the relative amount thereof. The invention further discloses a kit for detecting the allele. The kit comprises a plurality of sealed centrifuge tubes which are respectively filled with a reaction liquid 1, a reaction liquid 2, a reaction liquid 3, an EZ Taq enzyme mixed liquid, a standard liquid 1 and a standard liquid 2 as well as the kit which separates and packages the centrifuge tubes in a centralized manner. The kit and the detection method provided by the invention are simple and convenient to operate, short in time consumed, strong in specificity and high in sensitivity. The kit and the detection device can be widely applied to detecting the HLA-B*58:01 allele and clinically avoiding severe untoward effects of skins of the HLA-B*58:01 allele patients caused by using allopurinol.

Description

Technical field [0001] The present invention relates to a method for detecting HLA-B*58:01 alleles, and a kit used for the method, in particular to the use of real-time quantitative fluorescent polymerase chain reaction technology to detect HLA-B*58:01 alleles And apply the test results to clinical methods and kits to guide the use of allopurine drugs. Background technique [0002] Adverse drug reactions (adverse drug reactions, referred to as ADR), in accordance with the provisions of the WHO International Drug Surveillance Cooperative Center, refer to normal doses of drugs used to prevent, diagnose, treat diseases or regulate physiological functions that are harmful and have nothing to do with the purpose of medication Reaction. Adverse drug reactions are huge hidden dangers in the treatment of patients. According to statistics, the total number of people who die from irrational drug use in the world reaches 19 million every year. 2.2 million people in the U.S. experience si...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q1/6858C12Q1/686C12Q2600/106C12Q2600/156C12Q2531/113C12Q2561/101C12Q2561/113
Inventor 刘志文肖伟明阙秋华孙祖勇
Owner GUANGDONG UNITY BIOTECH
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