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Detection kit for human fructosediphosphate aldolase B gene and detection method thereof

A diphosphate aldolase and detection kit technology, applied in the field of genetic engineering, can solve the problems of low throughput, complicated operation, and long time consumption, and achieve the effects of less time, reducing false positives, and avoiding subjectivity

Active Publication Date: 2017-06-27
GUANGDONG HUAMEI ZHONGYUAN BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the main technology for detecting the human fructose diphosphate aldolase B gene is gene sequencing, which has the disadvantages of low throughput, complicated operation, and long time consumption.

Method used

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  • Detection kit for human fructosediphosphate aldolase B gene and detection method thereof
  • Detection kit for human fructosediphosphate aldolase B gene and detection method thereof
  • Detection kit for human fructosediphosphate aldolase B gene and detection method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Example 1: Composition and method of use of human fructose diphosphate aldolase B gene detection kit

[0041] The detection kit includes Taqman probes for specifically recognizing three polymorphic sites on the human fructose bisphosphate aldolase B gene and for amplifying the corresponding polymorphic site gene fragments. Primer pairs, specifically as shown in the table below:

[0042] Table 3. Composition of the detection kit

[0043]

[0044] The using method of described human fructose diphosphate aldolase B gene detection kit:

[0045] 1. Extraction of sample DNA

[0046] According to the actual situation of the sample, choose the appropriate method to extract the sample DNA, common methods include magnetic bead method, anion exchange resin method and alkaline lysis method, etc.;

[0047] 2. Fluorescent quantitative PCR reaction

[0048] The amplification system of the fluorescent quantitative PCR reaction is as follows:

[0049] Table 4. Real-time quantita...

Embodiment 2

[0056] Embodiment 2: the establishment of standard collection of illustrative plates:

[0057] According to the usage method of embodiment 1, the homozygous standard substance and the heterozygous standard substance in the human fructose diphosphate aldolase B gene detection kit are detected, obtain as follows figure 1 and 2 The detection profile of the homozygous standard is shown as well as image 3 Detection profiles of heterozygous standards are indicated.

Embodiment 3

[0058] Embodiment 3: No. 1 sample detection

[0059] According to the using method of embodiment 1, No. 1 sample is detected, obtain as follows Figure 4 As shown in the graph, it can be seen that sample No. 1 is a homozygous wild type.

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Abstract

The invention provides a detection kit for a human fructosediphosphate aldolase B gene and a detection method thereof. The detection kit comprises a Taqman probe which is used for specifically recognizing three polymorphic sites on the human fructosediphosphate aldolase B gene and a primer pair which is used for amplifying gene segments corresponding to the polymorphic sites, wherein the three polymorphic sites include A150P, A175D and N335K; the probe includes a wild type probe and a mutant type probe; and the primer pair includes an upstream primer and a downstream primer. The kit provided by the invention can quickly and efficiently detect the genotypes at the three polymorphic sites on the human fructosediphosphate aldolase B gene by performing sextuple fluorogenic quantitative PCR reaction on a sample and is a quick, convenient, economical and efficient hereditary fructose intolerance gene detection kit.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a human fructose diphosphate aldolase B gene detection kit and a detection method thereof. Background technique [0002] Hereditary Fructose Intolerance (HFI) is a kind of fructose metabolism disease. It is an autosomal recessive genetic disease. Deficiency or reduced activity, resulting in abnormal fructose metabolism in patients. [0003] At present, the main technology for detecting human fructose diphosphate aldolase B gene is gene sequencing, which has the disadvantages of low throughput, complicated operation, and long time consumption. [0004] Fluorescent quantitative PCR technology is a relatively mature nucleic acid detection method. Its advantages are: easy operation, less pollution due to closed-tube operation, and fast result analysis. Among them, the Taqman probe technology is a gene detection technology for distinguishing allele types by detecting th...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6883C12Q2600/156C12Q2563/107C12Q2545/114C12Q2537/143C12Q2561/101
Inventor 黄源坚郑文彦高静杜蔚安王邦超彭百华吴智锋周玉
Owner GUANGDONG HUAMEI ZHONGYUAN BIOLOGICAL SCI & TECH
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