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Sudden cardiac death rapid gene detection kit and detection method

A technology for sudden cardiac death and genetic testing, applied in biochemical equipment and methods, microbiological measurement/inspection, etc., can solve problems such as increasing the risk of sporadic atrial fibrillation

Inactive Publication Date: 2016-09-07
GUANGZHOU INST OF SPORTS SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some subtle changes in the NUP155 gene may also increase the risk of common sporadic atrial fibrillation

Method used

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  • Sudden cardiac death rapid gene detection kit and detection method
  • Sudden cardiac death rapid gene detection kit and detection method
  • Sudden cardiac death rapid gene detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0130] Amplification reagents consist of components in the following volumes:

[0131]

[0132] The internal reference upstream primers are SEQ NO.1, SEQ NO.5, SEQ NO.9, SEQ NO.13, SEQ NO.17, SEQNO.21, SEQ NO.25, and the polymorphic primers are SEQ NO.2, SEQ NO.6, SEQ NO.10, SEQ NO.14, SEQ NO.18, SEQ NO.22, SEQ NO.26, SEQ NO.3, SEQ NO.7, SEQ NO.11, SEQ NO.15, SEQ NO.19, SEQNO.23, SEQ NO.27, downstream primers are SEQ NO.4, SEQ NO.8, SEQ NO.12, SEQ NO.16, SEQ NO.20, SEQ NO.24, SEQ NO. 28. The concentration of the internal reference upstream primer is 10uM, the concentration of the polymorphism primer is 10uM, and the concentration of the downstream primer is 10uM.

[0133] The PCR amplification procedure is as follows:

[0134]

Embodiment 2

[0136] Amplification reagents consist of components in the following volumes:

[0137]

Embodiment 3

[0139] Amplification reagents consist of components in the following volumes:

[0140]

[0141] In order to verify the above information, the amplification product can be detected and analyzed on the ABI3130 Genetic Analyzer to verify the above detection results of the present invention.

[0142] The loading mixture is composed of deionized formamide and molecular weight internal standard. Mix 10ul of the loading mixture with 1ul of the amplification product or allele analysis standard to avoid bubbles, denature at 95°C for 5min, perform electrophoresis, and use ABI3130 for genetic analysis. Instrument detection and analysis to verify the analysis results.

[0143] The amplification is performed by AS-PCR (allele-specific PCR), and polymorphism typing is detected by agarose gel electrophoresis.

[0144] The human genomic DNA detected is the DNA obtained by processing the source sample using the Chelex method, magnetic bead extraction method or phenol / chloroform extraction ...

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Abstract

The present invention discloses a rapid gene detection kit for sudden cardiac death. The kit comprises 7 SNP polymorphism detections corresponding to genes such as KCNE1, KCNH2, SCN5A, KCNJ11, NUP155, CACNA1C and CACNB2B. The present invention further discloses a rapid gene detection method for sudden cardiac death. According to the present invention, semi-nested AS-PCR amplification and agarose gel electrophoresis detection are performed, and genotyping is performed according to the DNA electrophoresis bands to screen the individual with the potential sudden cardiac death risk so as to achieve the purpose of sudden cardiac death prevention; and the method has characteristics of simple operation, short detection time, low cost, high sensitivity, strong specificity, good social benefits and broad application prospects.

Description

technical field [0001] The invention relates to the technical field of biological gene detection, in particular to a sudden cardiac death rapid gene detection kit and detection method. Background technique [0002] Sudden cardiac death (sudden cardiac death, SCD) refers to the sudden death of the heart that loses effective contraction due to cardiac electrical disorder or heart failure. In recent years, SCD cases have occurred frequently, and the number has been increasing year by year. At present, the routine clinical detection methods for SCD are mainly to screen family history and personal history, and at the same time to perform physical examination and electrocardiogram examination. . However, these detection methods are not highly targeted, the detection rate is low, and they have great limitations. [0003] Domestic and foreign studies on sudden cardiac death have shown that there are some gene mutations at the genome level in the deceased, most of which are SNP or...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156
Inventor 陈伟民娄婧婧李长杰
Owner GUANGZHOU INST OF SPORTS SCI
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