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Alzheimer disease early-stage lesion CR-1 gene mRNA level in-situ hybridization detection kit

An in situ hybridization and kit technology, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as human and environmental damage, and achieve the effects of convenient operation, strong specificity and high sensitivity

Inactive Publication Date: 2015-06-17
NATUREGEN BIOTECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although isotope markers have the advantages of high sensitivity and clear background, but because radioactive isotopes can cause harm to people and the environment, they have recently tended to be replaced by non-isotopes.

Method used

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  • Alzheimer disease early-stage lesion CR-1 gene mRNA level in-situ hybridization detection kit
  • Alzheimer disease early-stage lesion CR-1 gene mRNA level in-situ hybridization detection kit
  • Alzheimer disease early-stage lesion CR-1 gene mRNA level in-situ hybridization detection kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes, markers, and instructions designed with the CR-1 gene as the detection target gene, wherein:

[0058] Digoxigenin was selected as the probe label in this embodiment.

[0059] Kit preparation:

[0060] Digestive solution 100μL / tube 1 tube / box Colorless transparent liquid

[0061] Protective solution 100μL / tube 1 tube / box Colorless transparent liquid

[0062] Pre-hybridization solution 1300μL / tube 2 tubes / box Colorless transparent liquid

[0063] Sense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0064] Antisense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0065] Blocking solution 1000μL / tube 1 tube / box Colorless transparent liquid

[0066] Alkaline phosphatase antibody 1μL / tube 1 tube / box Colorless transparent liquid

[0067] Chromogen A 175μL / tube 1 tube / box Yellow liquid

...

Embodiment 2

[0082] The implementation process of the application of nucleic acid in situ hybridization detection method to the gene expression of patients with Alzheimer's disease:

[0083] 1).Take two specimens to be tested;

[0084] 2). Add 50 ml of digestive solution (100 μL of digestive solution plus 99.9 ml of 1× buffer Ⅰ, which is the concentration used) in a glass tank, preheat in a water bath at 37°C for 10 minutes, put 16 slides in, and treat at 37°C for 12 minutes , and then washed with 1× buffer I for 5 min;

[0085] 3). Use 0.2% protection solution (protection solution 1ml plus 1× buffer , 99ml is the concentration used), washed for 10 minutes, washed with three-distilled water for 5 minutes (the above process was carried out in a glass tank), took out the slide, and let it dry naturally;

[0086] 4). Put the slides into a humidifying box, add 25 μL / slice of pre-hybridization solution (add to the place where there are cells), cover with a cover glass, cover the humidifying ...

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Abstract

The invention discloses an in-situ hybridization detection kit, which comprises a hybridization probe and a marker, and further discloses a method utilizing the in-situ hybridization detection kit to detect the CR-1 gene mRNA, which is closely related with the Alzheimer disease (AD) early-stage pathologic evolution. The method comprises the following steps: (1) under a condition that a hybridization probe and a target sequence can form a stable hybridization complex, contacting RNA to be detected in a substrate with a hybridization probe so as to form a hybridization complex; (2) detecting the obtained hybridization complex. The provided kit and detection method can detect the expression function of CR-1 gene in the mRNA level, can detect the cancer in an earlier stage compared with the medical imaging technology and conventional clinical biochemical index detection, can achieve real mRNA level screening in the early-stage lesion of AD so as to prevent and treat AD in advance, moreover, have the advantages of simpleness, convenience, and low cost, and thus are convenient to promote and use in hospitals.

Description

technical field [0001] The present invention relates to the field of biological detection, and more specifically relates to detection technology related to the change of mRNA expression (pathological evolution process) in the pre-pathological stage of senile dementia (AD). Background technique [0002] Alzheimer's disease (AD) is the most common type of dementia, accounting for 60%-80% of all dementias, and it is also one of the diseases with high disability rate and heavy burden. By 2006, the death rate of heart disease, the number one killer of human beings, had dropped by 11.5% compared with 2000, and the death rate of stroke, the second killer, had dropped by 18.1%. 5th cause of death. With the increase in global population size and life expectancy, Alzheimer's disease has become a world health problem. According to future forecasts, the prevalence of dementia will nearly double every two decades, and will reach 65.7 million by 2030. In 2050 it will reach 115.4 million...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/118C12Q2600/158
Inventor 张玉丽裘建英张云福裘霖
Owner NATUREGEN BIOTECH SHANGHAI
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