Genetic diagnosis kit of Alzheimer's disease

A technology for Alzheimer's disease and genetic diagnosis, applied in the field of molecular biology, to achieve rapid results

Pending Publication Date: 2016-09-28
江苏雄鸣医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are no patents or reports on the molecular diagnosis of AD diseases by this technology. The establishment of this patent method provides technical support for the rapid diagnosis and early screening of AD.

Method used

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  • Genetic diagnosis kit of Alzheimer's disease
  • Genetic diagnosis kit of Alzheimer's disease
  • Genetic diagnosis kit of Alzheimer's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Gene determination, primer design, kit composition and amplification procedure

[0049] The 10 genes involved in the present invention are related to AD disease, and their correlation has been reported in the literature. The sequences of 10 genes can be obtained by the following methods.

[0050] On the website http: / / www.ncbi.nlm.nih.gov / , the gene sequence can be retrieved according to the following gene ID.

[0051] Table 1 Gene ID

[0052]

[0053] In the primer design process, the design is mainly based on the conserved regions of each gene, and the primers are used to specifically amplify all the CDS regions of each gene. The forward and reverse primers are designed in the 5'-UTR and 3'-UTR regions, respectively, to ensure that the amplified product can cover all the CDS regions, enabling the kit to detect mutation sites on all CDS regions of each disease-causing gene.

[0054] Taking the CLU gene as an example, the mRNA sequence is shown below, where the underl...

Embodiment 2

[0065] Example 2 Genetic testing of Alzheimer's disease

[0066] According to the kit of embodiment 1, the application of embodiment 2 is carried out, and the specific steps are as follows:

[0067] In the genetic diagnosis kit for Alzheimer's disease provided by the present invention, the sample to be tested can be blood, body fluid, hair, bone, cell or tissue specimens of the individual to be tested. In this example, the standard cell line as in Example 1 was used.

[0068] 1. Extraction of RNA from human fibroblast skin cells to be tested:

[0069] 1. Take the sample to be tested, add 1 ml Trizol solution and mix on ice, mash the homogenate; let it stand on ice for 10 min;

[0070] 2. Add 200 μl of chloroform to the homogenate, shake vigorously for 15s, and incubate at room temperature for 10 min

[0071] 3. Centrifuge at 4℃, 12000 g / min, l for 15 min

[0072] 4. Take the water phase (upper layer), add 500 μl isopropanol, shake gently, and incubate at room temperature for 10-15 min

[...

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Abstract

The invention discloses a genetic diagnosis kit of Alzheimer's disease. The kit can detect, research and report 10 virulence genes related to Alzheimer's disease, including APP, APOE, PSEN1, PSEN2, GSK3beta, DYRK1A, Tomm40, CLU, PICALM and TAU. A detecting method comprises the following steps: designing and synthesizing specific primers of all genes, collecting the sample of a individual to be detected, adopting the RT-PCR technology to carry out sequencing analysis on obtained specificity DNA products of all genes, comparing the sample sequence with the normal gene sequence, and analyzing whether pathogenic mutations exist to evaluate the morbidity risk of the individual. The genetic diagnosis kit of Alzheimer's disease can detect different pathogenic mutation sites of CDS at a time and has the advantages of high simpleness, convenience, rapidity and sensitivity, reliable preparation and excellent specificity.

Description

Technical field [0001] The invention relates to a genetic diagnosis kit for genetic diseases, more specifically a genetic diagnosis kit for Alzheimer's disease (also known as Alzheimer's disease), which belongs to the technical field of molecular biology. Background technique [0002] Alzheimer’s disease (AD), also known as Alzheimer’s disease, is a neurodegenerative disease that often occurs in the elderly. The disease was first recorded and described by German physician Alois Alzheimer in 1906. It is characterized by (1) its characteristic pathological changes are β-amyloid protein deposition and Tau protein hyperphosphorylation to form neurofibrillary tangles in nerve cells. Loss and glial cell proliferation are neurodegenerative diseases. The disease manifests as mild memory impairment in the early stage. With the development of the disease, the patient’s cognitive abilities (such as memory, calculation, thinking, judgment, etc.) gradually decline, intelligence declines, em...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/156
Inventor 陈实黎寒
Owner 江苏雄鸣医药科技有限公司
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