Kit for examining Liddle's related gene mutation

A kit and gene technology, applied in the field of molecular biology, can solve the problems of low sensitivity and specificity, strict detection time requirements, unbearable cost, etc., to avoid uncertain primers, improve detection accuracy and stability, increase Effects on Clinical Detection Rate and Accuracy

Inactive Publication Date: 2014-10-08
百世诺(北京)医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] 1. There are many types of inspections;
[0011] 2. Low sensitivity and specificity;
[0012] 3. Susceptible to factors such as drugs and mental conditions;
[0013] 4. Strict detection time requirements;
[0023] However, the cost of directly using a large-scale DNA parallel sequencing platform is still unaffordable for individuals

Method used

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  • Kit for examining Liddle's related gene mutation
  • Kit for examining Liddle's related gene mutation
  • Kit for examining Liddle's related gene mutation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] 1. Probe design:

[0048] Synthesize streptomycin-labeled probes according to the table. The probe solution uses the SureSelect buffer system of agilent company.

[0049] 2. Genome extraction:

[0050] Qiagen FlexiGene DNA Kit (Code No: 51204) was used to extract the genomes of 96 samples to be tested, OD 260 / 280 When the value reaches 1.8-2.0, take 1-3g each as the starting template.

[0051] 3. Sample preparation before sequencing

[0052] 1. Target gene fragmentation:

[0053] Take the quantified 270 copies of genomic DNA and dilute to 20-35ng / L. Take 130L and fragment them separately with a sonicator.

[0054] 2. AMpure XP fragment selection

[0055] 1. Take a 96-well plate, add 80-100L magnetic beads, then add sample DNA, pipette repeatedly to mix;

[0056] 2. Place the mixture at 26°C for 5 minutes;

[0057] 3. Put the 96-well plate on the magnetic plate for 3 minutes;

[0058] 4. Carefully pipette all the supernatant into a new well, be careful not to t...

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Abstract

The invention relates to a kit made for detecting Liddle's syndrome related gene mutation samples, and concretely relates to a product for detecting Liddle's syndrome related gene by adopting a large scale parallel sequencing platform technology. A detection method comprises the following steps: 1, uniquely designing and making a capture probe against all exon fragments of gene SCNN1B and SCNN1G; 2, designing a unique joint with a labeling sequence; 3, carrying out PCR amplification on the sequence of the probe by using a general primer; and 4, designing a unique capture operation of mixed target fragments. The method and the kit have the advantages of large flux of prepared large scale parallel sequencing platform samples, high efficiency, simple operation, and great reduction of the sequencing examination cost.

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 detecting Liddle's-related gene mutations, and is a kit for Liddle's-related gene mutations applied to the next-generation sequencing platform technology. technical background [0002] Liddle's syndrome is an autosomal dominant monogenic hypertension characterized clinically by early-onset hypertension, hypokalemia, hyporenin, hypoaldosteronemia, and response to epithelial sodium channel (ENaC) inhibitors Amiloride or triamterene is sensitive, but insensitive to aldosterone receptor antagonist spironolactone, and the effect of conventional hypertension drug treatment is not good. [0003] Liddle's syndrome is a rare disorder, but its incidence may be higher than expected, and it is one of the most common causes of monogenic hypertension. The disease has been reported in different races, including Caucasians...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2535/122C12Q2525/191C12Q2531/113
Inventor 刘哲
Owner 百世诺(北京)医疗科技有限公司
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