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Primer probe for diagnosing corneal dystrophy caused by human TGF beta I gene 555 locus mutation and detection method

A dystrophy, primer probe technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as Thiel-Behnke corneal dystrophy, achieve high annealing Tm value, primers High matching rate and the effect of increasing the signal-to-noise ratio

Inactive Publication Date: 2018-03-16
奥斯汀生命科学技术公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Type Q555TBCD, causing Thiel-Behnke corneal dystrophy

Method used

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  • Primer probe for diagnosing corneal dystrophy caused by human TGF beta I gene 555 locus mutation and detection method
  • Primer probe for diagnosing corneal dystrophy caused by human TGF beta I gene 555 locus mutation and detection method
  • Primer probe for diagnosing corneal dystrophy caused by human TGF beta I gene 555 locus mutation and detection method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] A primer probe for diagnosing corneal dystrophy caused by mutation of human TGFβI gene 555, comprising the following sequence:

[0058] The specific primer pair for amplifying the 555 site of the TGFβI gene, the base sequence of the specific primer pair is as follows:

[0059] F: 5'-GGAGTCTACACAGTCTTTGCTC-3';

[0060] R: 5'-CAAGTCAATCCTTGATGTGCC-3';

[0061] For the specific wild-type Taqman fluorescent probe of the wild-type site R555, the base sequence of the fluorescent probe is as follows:

[0062] R555: 5'-FAM-CCAAGAGAACGGAGCAGAC-MGB-3';

[0063] For the specific mutant Taqman fluorescent probe of mutant site W555, the base sequence of the fluorescent probe is as follows:

[0064] W555: 5'-JOE-CCAAGAGAATGGAGCAGAC-MGB-3'.

Embodiment 2

[0066] A primer probe for diagnosing corneal dystrophy caused by mutation of human TGFβI gene 555, comprising the following sequence:

[0067] The specific primer pair for amplifying the 555 site of the TGFβI gene, the base sequence of the specific primer pair is as follows:

[0068] F: 5'-GGAGTCTACACAGTCTTTGCTC-3';

[0069] R: 5'-CAAGTCAATCCTTGATGTGCC-3';

[0070] For the specific wild-type Taqman fluorescent probe of the wild-type site R555, the base sequence of the fluorescent probe is as follows:

[0071] R555: 5'-FAM-CCAAGAGAACGGAGCAGAC-MGB-3';

[0072] For the specific mutant Taqman fluorescent probe at the mutant site Q555, the base sequence of the fluorescent probe is as follows:

[0073] Q555: 5'-CY5-GTACACGGACCTCACGGAG-MGB-3'.

Embodiment 3

[0075] A primer probe for diagnosing corneal dystrophy caused by mutation of human TGFβI gene 555, comprising the following sequence:

[0076] The specific primer pair for amplifying the 555 site of the TGFβI gene, the base sequence of the specific primer pair is as follows:

[0077] F: 5'-GGAGTCTACACAGTCTTTGCTC-3';

[0078] R: 5'-CAAGTCAATCCTTGATGTGCC-3';

[0079] For the specific wild-type Taqman fluorescent probe of the wild-type site R555, the base sequence of the fluorescent probe is as follows:

[0080] R555: 5'-FAM-CCAAGAGAACGGAGCAGAC-MGB-3';

[0081] Two specific mutant Taqman fluorescent probes for mutant sites W555 and Q555 respectively, the base sequences of the fluorescent probes are as follows:

[0082] W555: 5'-JOE-CCAAGAGAATGGAGCAGAC-MGB-3';

[0083] Q555: 5'-ROX-CCAAGAGAACAGAGCAGAC-MGB-3'.

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Abstract

The present invention provides a primer probe for diagnosing corneal dystrophy caused by variation of human TGFβI gene 555 site, including specific primer designed for TGFβI gene 555 site, specific wild-type Taqman for wild-type site R555 Fluorescent probes, and at least one of the two specific mutant Taqman fluorescent probes for mutant sites W555 and Q555, each probe is connected to a different fluorescent reporter group, so as to separate monitoring, and does not require the addition of internal references. The present invention also provides a detection method for human TGFβI gene 555 site variation based on the above-mentioned primer probe, which has the advantages of high sensitivity, strong specificity, non-pollution, good accuracy, convenience and quickness, and is suitable for application in clinical case analysis and testing at work.

Description

technical field [0001] The invention belongs to the technical field of gene site mutation detection, in particular to a primer probe and a detection method for diagnosing corneal dystrophy caused by the 555 site mutation of human TGFβI gene. Background technique [0002] Corneal dystrophy is a group of symmetrical and hereditary diseases of both eyes. Under the induction of certain genetic abnormalities, proteins in different layers of the cornea aggregate atypically, and deposits are gradually formed to make the cornea turbid and lose its transparency. Severe Affect the vision and quality of life of patients. Human transforming growth factor-beta induce (TGFβI) gene, also known as βIGH3 gene, is the first confirmed corneal dystrophy gene, more than 50% of human corneal dystrophy is caused by certain mutations in BIGH3 . At present, the clinical method for the treatment of myopia is excimer laser therapeutic keratectomy PTK (Photo Therapeutic Keratectomy), which is mainly ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6851C12N15/11
CPCC12Q1/6883C12Q1/6851C12Q2600/156C12Q2531/113C12Q2563/107
Inventor 王瀚生
Owner 奥斯汀生命科学技术公司
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