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In-situ hybridization detection kit for microRNA-182 level in early stage of pathological evolution of various cancers, and detection method and application thereof

A detection kit and in situ hybridization technology, applied in the field of related detection technology, can solve the problems of non-decreasing mortality, drug resistance of tumor cells, failure of anti-cancer war, etc., and achieve convenient operation, high sensitivity and strong specificity Effect

Inactive Publication Date: 2012-05-09
SUZHOU FUYING GENE TECH
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  • Summary
  • 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.
[0016] 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

Method used

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  • In-situ hybridization detection kit for microRNA-182 level in early stage of pathological evolution of various cancers, and detection method and application thereof
  • In-situ hybridization detection kit for microRNA-182 level in early stage of pathological evolution of various cancers, and detection method and application thereof
  • In-situ hybridization detection kit for microRNA-182 level in early stage of pathological evolution of various cancers, and detection method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0052] The in situ hybridization kit of this example was prepared according to conventional methods, and the kit included a hybridization probe designed with MICRORNA-182, a label, and instructions, wherein: the probe label of this example was digoxin.

[0053] Kit hybridization solution composition:

[0054] 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 Chr...

Embodiment 2

[0062] The implementation process of applying nucleic acid in situ hybridization detection method to the expression level of MICRORNA-182 in blood samples of each group:

[0063] 1).Take two specimens to be tested;

[0064] 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;

[0065]3). Wash with 0.2% protection solution (protection solution 1ml plus 1× buffer Ⅰ, 99ml is the used concentration) for 10 minutes, three-distilled water for 5 minutes (the above process is carried out in a glass tank), take out the slide and let it naturally dry;

[0066] 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...

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Abstract

The invention discloses an in-situ hybridization detection kit which comprises a hybridization probe and markers. The invention also discloses a method for applying the kit in in-situ hybridization detection of micro RNA-182 having a close relationship with pathological evolution of various cancers in an early stage, and the method comprises the following steps: (1) under the condition that the hybridization probe and a target sequence can form a stable hybrid complex, contacting RNA to be detected in a substrate with the hybridization probe so as to form a hybrid complex; (2) detecting the hybrid complex. The kit and the detection method provided in the invention can detect the expression level of micro RNA-182 at RNA level, enable detection indexes to be obtained at an earlier stage than medical imaging and conventional clinical biochemical detection do, and can realize genuine precancerous lesion RNA level screening; meanwhile, the detection method is simple and convenient, costs little and is convenient for popularization and application in county hospitals.

Description

technical field [0001] The present invention relates to the field of biological detection, more specifically, relates to the relevant detection technology related to the change of RNA expression (pathological evolution process) of various cancer pathological evolutions. 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 (may be higher in poor areas, and 200,000 yuan in developed areas). For more than 7 million patients, the annual cost is 1.4 trillion y...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 裘建英张云福张玉丽裘霖
Owner SUZHOU FUYING GENE TECH
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