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Primer combination, probe, gene chip, kit and system for detecting HLA-B5801 gene

A primer combination and gene chip technology, which can be used in DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., and can solve problems such as adverse reactions

Pending Publication Date: 2021-09-14
国家卫生健康委科学技术研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, allopurinol is prone to cause various forms of adverse drug reactions, ranging from mild macular MPE to severe adverse drug reactions, even including hypersensitivity syndrome HSS, syndrome SJS, and toxic epidermal necrosis syndrome, etc.

Method used

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  • Primer combination, probe, gene chip, kit and system for detecting HLA-B5801 gene
  • Primer combination, probe, gene chip, kit and system for detecting HLA-B5801 gene
  • Primer combination, probe, gene chip, kit and system for detecting HLA-B5801 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1 Primer, probe and test kit

[0064] 1. Primer combination

[0065] According to the sequence information of the target gene HLA-B5801 allele, primers were designed, and the designed primers were evaluated for secondary structure and Tm value, and finally obtained the specificity, high sensitivity, and can be detected under the same reaction conditions. A primer of the nucleotide sequence shown in SEQ ID NO.1-14 of the present invention is used to determine whether the biological sample to be tested is suitable for allopurinol.

[0066] Secondary structure evaluation and Tm value evaluation can be carried out by any method commonly used in the art. For example, in one embodiment of the present invention, DNA folding form is used to evaluate its secondary structure. For details, see (http: / / unafold. rna.albany.edu / ?q=mfold / DNA-Folding-Form), and then use MLPA's own software RaW-Probe to evaluate its Tm value.

[0067] It should be noted that the above-menti...

Embodiment 2

[0079] The detection system of embodiment 2 HLA-B5801 gene

[0080] Such as figure 1 As shown, the system for the rapid screening of biological samples prone to recurrent miscarriage provided by the invention comprises:

[0081] The whole genome DNA extraction unit 101 is used to extract the whole genome DNA sample in the biological sample; the whole genome DNA sample processing unit 102 uses a primer combination or a Taqman probe combination or a kit to perform fluorescent quantitative PCR with the whole genome DNA sample; The sample analysis unit 103 is used to detect the fluorescence signal of the amplification product, analyze the typing result of the sample according to the amplification curve generated by the fluorescence signal, and give the judgment result of whether the sample can use allopurinol.

[0082] The features and advantages described above in the method for screening biological samples susceptible to recurrent miscarriage are also applicable to the system f...

Embodiment 3

[0083] Screening and detection of embodiment 3 biological samples

[0084] 1. Sample acquisition and processing

[0085] Peripheral blood was collected by conventional methods. The peripheral blood is an isolated biological sample taken from the human body, and the whole genome DNA in the sample was extracted using a conventional DNA extraction kit to obtain a whole genome DNA sample. ThermoScientific NanoDrop 2000 UV spectrophotometer was used Measure the DNA sample concentration with a meter and dilute it to 20ng / μl for later use;

[0086] Of course, the biological sample can also use any other isolated biological sample capable of extracting whole genome DNA, such as amniotic fluid cells, skin tissue, hair, saliva, and muscle.

[0087] 2. Preparation of reaction solution

[0088] Dilute each probe in the kit to 500fmol / ul separately to prepare Probe Mix; then configure the reaction solution, namely

[0089]

[0090] 3. Fluorescent quantitative PCR reaction

[0091] T...

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Abstract

The invention provides a primer combination, a probe, a gene chip, a kit and a system for detecting an HLA-5801 gene, and relates to the technical field of biology. The primer combination, the probe, the gene chip and the kit T have a nucleotide sequence as shown in any one of SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.5, SEQ ID NO.6, SEQ ID NO.9 and SEQ ID NO.10. The primer combination can accurately detect applicability of allopurinol of a biological sample, and has good specificity, sensitive reaction and short detection time. The method is simple and has low cost and high clinical application value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer combination, probe, gene chip, kit and system for detecting HLA-B5801 gene. Background technique [0002] Human leukocyte antigen (HLA) is a group of genes located on chromosome 6. It is the gene complex with the highest allelic polymorphism among known genes so far. More than 2,700 alleles have been found in the world, including HLA- B5801 allele. HLA-B5801 is a gene in the huge human genome. According to current research, it is highly related to severe skin adverse reactions caused by drug allergy, including allopurinol. In the Asian population (including my country), the positive rate of HLA-B5801 is relatively high, which has become one of the important concerns of the Asian population taking allopurinol. [0003] Allopurinol is a xanthine oxidase inhibitor that can effectively reduce blood uric acid. It is a common prescription drug for the treatment of gout, acute ur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6881C12Q1/6858C12N15/11
CPCC12Q1/6881C12Q1/6858C12Q2600/106C12Q2600/156C12Q2531/113C12Q2561/101C12Q2563/107
Inventor 王彬彬王晶
Owner 国家卫生健康委科学技术研究所
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