In situ hybridization detection kit for mRNA level at precancerous stage of breast cancer, detection method and application thereof

A detection kit and in situ hybridization technology are applied in the field of detection technology related to mRNA expression changes, which can solve the problems of non-decreased mortality, drug resistance of tumor cells, failure of anti-cancer battles, etc., and achieve convenient operation, high sensitivity, specific effect

Active Publication Date: 2012-07-04
NATUREGEN BIOTECH SHANGHAI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] In 2005, eight institutions including the US Institutes of Health, the Cancer Institute, and the Centers for Disease Control and Prevention made an annual report, reviewing the anti-cancer war launched in 1972. The report believed that human beings had failed in the anti-cancer war, and the conclusion was that cancer died The rate did not decrease, and it listed several factors that caused the failure of the anti-cancer war: 1. Tumor cell heterogeneity (polymorphism); 2. Tumor cell drug resistance; 3. Incomplete design of anti-cancer drugs (animals) unscientific model design), etc.
[0015] In view of the fact that the current clinical diagnosis of cancer (imaging medicine and biochemical indicators are all diagnosed after tumor formation) is a late diagnosis, the treatment is also a late treatment, leading to a treatment model that does not reduce the mortality rate

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  • In situ hybridization detection kit for mRNA level at precancerous stage of breast cancer, detection method and application thereof
  • In situ hybridization detection kit for mRNA level at precancerous stage of breast cancer, detection method and application thereof
  • In situ hybridization detection kit for mRNA level at precancerous stage of breast cancer, detection method and application thereof

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Experimental program
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Embodiment 1

[0056] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes, markers, and instructions designed with the ZNF703 gene as the detection target gene, wherein:

[0057] Digoxigenin was selected as the probe label in this embodiment.

[0058] Kit hybridization solution composition:

[0059] 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 Chromoge...

Embodiment 2

[0067] The implementation process of applying the nucleic acid in situ hybridization detection method to the ZNF703 gene expression of each group of blood samples:

[0068] 1).Take two specimens to be tested;

[0069] 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;

[0070] 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;

[0071] 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 ti...

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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 in situ hybridization detection of an mRNA level of a zinc finger protein 703 (ZNF703) gene closely related with pathological evolution at a precancerous stage of breast cancer by using the kit. The method comprises the following steps that: (1) under a condition that the hybridization probe and a target sequence can form a stable hybrid complex, the RNA requiring detection in the substrate contacts the hybridization probe to form a hybrid complex; and (2) the hybrid complex is detected. With the kit and the detection method of the present invention, the expression amount of the ZNF703 gene on the mRNA level can be detected, such that the provision time of the index is earlier than the provision time of the index by medical imaging or by existing clinical biochemical detection, the screening on the mRNA level at the precancerous stage can be achieved truly. In addition, the detection method of the present invention has characteristics of simpleness, convenience and low cost, and is applicable for popularization and application in county hospitals and district hospitals.

Description

technical field [0001] The invention relates to the field of biological detection, more specifically, relates to the relevant detection technology related to the mRNA expression change (pathological evolution process) of breast cancer premutation. Background technique [0002] According to the information provided by authoritative organizations at home and abroad, there are 2.6 million new cancer cases in my country every year, nearly 2.1 million deaths, and more than 7 million patients. Globally, there are 8 million new cancer patients every year, nearly 8 million deaths, and more than 8,400 patients. Ten thousand people, the number will double by 2020, this is a set of terrible figures. The cost of cancer diagnosis and treatment is getting higher and higher. The annual treatment cost of cancer patients is 200,000 yuan (poor areas may be higher, and developed areas may be much higher than 200,000 yuan), and there are more than 7 million patients. The annual cost is 1.4 trill...

Claims

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Application Information

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