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In-situ hybridization screening kit for mRNA (messenger Ribonucleic Acid) level of gene CD33 of prophase lesion of Alzheimer's disease (AD) and screening method and application thereof

An in situ hybridization and kit technology, applied in biochemical equipment and methods, microbial determination/inspection, 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: 2013-04-17
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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  • In-situ hybridization screening kit for mRNA (messenger Ribonucleic Acid) level of gene CD33 of prophase lesion of Alzheimer's disease (AD) and screening method and application thereof
  • In-situ hybridization screening kit for mRNA (messenger Ribonucleic Acid) level of gene CD33 of prophase lesion of Alzheimer's disease (AD) and screening method and application thereof
  • In-situ hybridization screening kit for mRNA (messenger Ribonucleic Acid) level of gene CD33 of prophase lesion of Alzheimer's disease (AD) and screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes, markers, and instructions designed with the CD33 gene as the detection target gene, wherein:

[0056] Digoxigenin was selected as the probe label in this embodiment.

[0057] digestive juice 100μL / tube 1 tube / box colorless transparent liquid protective fluid 100μL / tube 1 tube / box colorless transparent liquid Pre-hybridization solution 1300μL / tube 2 tubes / box colorless transparent liquid Right-sense hybridization solution 10μL / tube 1 tube / box colorless transparent liquid antisense hybridization solution 10μL / tube 1 tube / box colorless transparent liquid blocking solution 1000μL / tube 1 tube / box colorless transparent liquid Alkaline phosphatase antibody 1 μL / tube 1 tube / box colorless transparent liquid Chromogen A 175μL / tube 1 tube / box yellow liquid C...

Embodiment 2

[0065] The implementation process of CD33 gene expression in senile dementia patients by nucleic acid in situ hybridization detection method:

[0066] 1).Take two specimens to be tested;

[0067] 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;

[0068] 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;

[0069] 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 box tightly, and place in a...

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Abstract

The invention discloses an in-situ hybridization detection kit which comprises a hybridization probe and a marker. The invention further discloses a method for carrying out in-situ hybridization detection on the mRNA (messenger Ribonucleic Acid), which is closely related to the pathological evolution of the prophase of the Alzheimer's disease (AD), of a gene CD33 by using the kit, and the method comprises the following steps of: (1) enabling the RNA to be detected in a substrate to be in contact with the hybridization probe under the condition that a stable hybridization complex can be formed by the hybridization probe and a target sequence, thereby forming the hybridization complex; and (2) detecting the hybridization complex. According to the kit and the method, disclosed by the invention, the expression function of the gene CD33 can be detected at an mRNA level, the prophase earlier than an early phase detected by the existing clinical biochemical detection indicators and imaging medicine is detected, and the real mRNA level screening of the prophase lesion of the AD can be realized, thus the aim of preventive diagnosis and treatment is achieved. Meanwhile, the method disclosed by the invention is simple and convenient and is low in cost, thereby facilitating the popularization and application to 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
Inventor 张玉丽裘建英张云福裘霖
Owner NATUREGEN BIOTECH SHANGHAI
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