Differential diagnostic kit for myocardial hypertrophy genes

A technology for differential diagnosis and cardiac hypertrophy, applied in the field of molecular biology, can solve the problems of low efficiency, high cost of single nucleotide sequencing, and Sanger sequencing method cannot meet the requirements of sequencing, so as to reduce costs, improve detection accuracy and Stability, avoiding uncertainty about the effect of primers

Inactive Publication Date: 2018-05-15
FUWAI HOSPITAL CHINESE ACAD OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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Problems solved by technology

[0003] At present, Sanger sequencing, as a traditional sequencing technology, can only determine one base sequence in one reaction. Although it is accurate, the efficiency is low, and the cost of single nucleotide sequencing is high.
For the research of complex diseases with genetic heterogeneity, it is usually necessary to sequence the entire coding regions of several or even dozens of genes to find valuable mutation information, and Sanger sequencing cannot meet the requirements of sequencing

Method used

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  • Differential diagnostic kit for myocardial hypertrophy genes
  • Differential diagnostic kit for myocardial hypertrophy genes
  • Differential diagnostic kit for myocardial hypertrophy genes

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Embodiment Construction

[0033] 1. The present invention is described in further detail and completeness below by means of the examples, but the present invention is not limited to the scope of the examples.

[0034] 2. Use TIANamp Blood DNA Midi Kit (DP332) to extract the genome of the sample to be tested. The volume of elution buffer (TE) is not less than 200 μL, and the OD260 / 280 value reaches 1.8-2.0. Take 2 μL of 25ng / μL DNA as a starting point. start template.

[0035] 3. Quantify the sample with Qubit, and use ddH after quantification 2 O Dilute the g DNA sample to 25 ng / μL.

[0036] 4. Building a database

[0037] 5. DNA fragmentation of genomic samples and addition of adapters and tags

[0038] 6. Add 1.5ml of Ethanol to 4.5ml of TargetCap Stop Solution, and mix thoroughly to make the final concentration of Ethanol 25%. Bring 1X TargetCap Stop Solution to room temperature.

[0039] 7. Take out the Purification Beads and place them at room temperature for at least 30 minutes.

[0040] 8. P...

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Abstract

The invention discloses a differential diagnostic kit for myocardial hypertrophy genes. The differential diagnostic kit is characterized that the technology relates to the differential diagnostic kitfor detecting the myocardial hypertrophy genes by applying a large-scale parallel sequencing platform NGS (Next-Generation Sequencing) technique. The differential diagnostic kit is mainly characterized in that A, unique design and preparation of capture probes aiming at all exon fragments of genes such as ACTC1, ACTN2, ANKRD1, CALR3, CASQ2, CAV3, CSRP3, FHL1, JPH2, LDB3, MYBPC3, MYH6, MYH7, MYL2,MYL3, MYLK2, MYOZ2, MYPN, NEXN, PLN, PRKAG2, SLC25A4, TCAP, TNNC1, TNNI3, TNNT2, TPM1, TRIM63, TTN, VCL, GLA, TTR, LAMP2 and FLNC; B, unique design of a connector with a tag sequence; C, PCR (Polymerase Chain Reaction) amplification of the probe sequence by using a universal primer pair; D, capture operation of mixed target fragments with unique design.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medical diagnosis and biotechnology, relates to an in vitro diagnostic kit for cardiac hypertrophy gene mutation, and is a differential diagnosis kit for cardiac hypertrophy gene applied to the NGS technology of a new generation sequencing platform. Background technique [0002] Myocardial hypertrophy (MH) is a common pathological process of many cardiovascular diseases, such as valvular heart disease, cardiomyopathy, hypertension, etc. It is a compensatory response of myocardial tissue to cope with increased cardiac load , in the process of MH, increased protein synthesis, increased volume, myocardial fibrosis and interstitial components of cardiomyocytes are one of the independent risk factors for cardiovascular disease. MH can be divided into pathological hypertrophy and physiological hypertrophy. Physiological hypertrophy refers to myocardial hypertrophy caused by pregnancy, physic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 宋雷王继征惠汝太邹玉宝张禅那
Owner FUWAI HOSPITAL CHINESE ACAD OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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