A kit for detecting the mutation site of susceptibility gene for granular corneal dystrophy

A site and gene technology, applied in the field of kits for detection of mutation sites of susceptibility genes for granular corneal dystrophy, can solve the problems of long detection time, prone to false positives, and high cost, so as to reduce detection cost and shorten detection time time, the effect of rapid detection

Active Publication Date: 2021-08-10
BGI GENOMICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the MassARRAY mass spectrometry genotyping system can realize the detection of multiple sites in one reaction at the same time by means of multiplex PCR technology, for these 4 sites, due to the high similarity of the base sequences in the regions where the sites are located, multiple The specificity of PCR amplification primers is poor, false positives are prone to occur, and the false positives are high. For the detection of the above four sites, reaction wells need to be set separately for reaction, and the detection time is long and the cost is high.

Method used

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  • A kit for detecting the mutation site of susceptibility gene for granular corneal dystrophy
  • A kit for detecting the mutation site of susceptibility gene for granular corneal dystrophy
  • A kit for detecting the mutation site of susceptibility gene for granular corneal dystrophy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0136] Embodiment 1, be used to detect the preparation of the primer set of SNP site in TGFβI gene

[0137] The set of primers used to detect SNP sites in the TGFβI gene consists of primer pairs named Primer Pair A, Primer Pair B, and Primer Pair C, and extension primers named Extension Primer A1, Extension Primer A2, Extension Primer B, and Extension Primer C. Primer composition;

[0138] The primer pair A is composed of single-stranded DNAs named A-F and A-R respectively, and A-F and A-R are the single-stranded DNAs shown in sequence 1 and sequence 2 in the sequence listing, respectively;

[0139] Primer pair B is composed of single-stranded DNA named B-F and B-R respectively, and B-F and B-R are respectively the single-stranded DNA shown in sequence 3 and sequence 4 in the sequence listing;

[0140] Primer pair C is composed of single-stranded DNA named C-F and C-R respectively, and C-F and C-R are respectively the single-stranded DNA shown in sequence 5 and sequence 6 in ...

Embodiment 2

[0147] Example 2. Detection of TGFβI gene SNP sites——rs121909208, rs121909209, rs121909211 and rs121909215

[0148] Utilizing the set of primers in Example 1, using Agena Bioscience Inc.'s MassARRAY time-of-flight mass spectrometry biochip system and iPLEX Gold reagent (Agena Bioscience Inc.) to carry out genetic analysis on rs121909208, rs121909209, rs121909211 and rs121909215 of the blood samples of three people selected Typing detection, and preparation of these four sites for non-wild type (ie mutant) recombinant plasmids for verification. Among them, the nucleotides of the four SNP sites of the TGFβI gene of these three people were all detected and determined by the existing technology, and rs121909208, rs121909209, rs121909211 and rs121909215 were wild-type C, G, G and G, respectively. The experiment was repeated three times, and the specific steps are as follows:

[0149] 1) Genomic DNA extraction and sample preparation:

[0150] Extract genomic DNA from blood samples...

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Abstract

The invention discloses a kit for detecting the mutation site of susceptibility gene of granular corneal dystrophy. The kit of the present invention includes primer pair A, primer pair B, primer pair C, and extension primer A1, extension primer A2, extension primer B and extension primer C shown in Sequence 7, Sequence 8, Sequence 9 and Sequence 10; A consists of the single-stranded DNA shown in the 11-30th position of sequence 1 and the 11-30th position of sequence 2, and the primer pair B is represented by the 11-30th position of sequence 3 and the 11-30th position of sequence 4 The primer pair C is composed of the single-stranded DNA shown in the 11-30th position of sequence 5 and the 11-30th position of sequence 6. The kit of the invention can detect the nucleotides of rs121909208, rs121909209, rs121909211 and rs121909215, and can also be used for auxiliary diagnosis of granular corneal dystrophy.

Description

technical field [0001] The invention relates to a kit for detecting a mutation site of a susceptibility gene for granular corneal dystrophy in the field of biotechnology. Background technique [0002] Hereditary granular corneal dystrophy (Granular Corneal Dystrophy, GCD) is a general term for a series of primary progressive corneal lesions related to family inheritance. Most of the disease is an autosomal dominant genetic disease, originating in the cornea, rarely accompanied by other eye diseases or systemic diseases; the onset is mostly before the age of 20; the course of the disease is slow, and there is no new blood vessel growth in the lesion area; it only invades the cornea at the beginning A certain layer of the cornea can spread to adjacent layers in the late stage, and even affect the whole thickness of the cornea; drug treatment is ineffective. [0003] So far, a total of 12 genes related to GCD have been reported, among which the TGFβI (transforming growth facto...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2535/125C12Q2565/627
Inventor 郑建超张燕张春杨王夏曼张红云
Owner BGI GENOMICS CO LTD
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