MicroRNA-155 level in various types of cancerous lesions prophase in situ hybridization detection kit, detection method and application thereof

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

Inactive Publication Date: 2012-08-08
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.
[0014] In view of the current clinical diagnosis of cancer (imaging medicine and biochemical indicators are all diagnosed after tumor formation) is a late diagnosis, treatment is also a late treatment, leading to a treatment model that does not reduce mortality

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  • MicroRNA-155 level in various types of cancerous lesions prophase in situ hybridization detection kit, detection method and application thereof
  • MicroRNA-155 level in various types of cancerous lesions prophase in situ hybridization detection kit, detection method and application thereof
  • MicroRNA-155 level in various types of cancerous lesions prophase in situ hybridization detection kit, detection method and application thereof

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Embodiment 1

[0052] The in situ hybridization kit of this embodiment was prepared according to conventional methods, and the kit included hybridization probes, markers, and instructions designed with microRNA-155 as the target gene for detection, wherein: the probe markers of this embodiment were chosen to be high pungent.

[0053] 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 liqui...

Embodiment 2

[0061] The implementation process of applying the nucleic acid in situ hybridization detection method to the microRNA-155 expression level of each group of blood samples:

[0062] 1).Take two specimens to be tested;

[0063] 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;

[0064] 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;

[0065] 4). Put the slides into the humidity box, add 25 μL / slice of pre-hybridization solution (add to the place where there are cells), cover with a cover glass, cover the humidity bo...

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Abstract

The invention discloses an in situ hybridization detection kit, which includes a hybridization probe and a marker. The invention also discloses an in situ hybridization detection method of microRNA-155 which is closely related to cancer prophase pathologic development by the use of the kit and comprises the following steps of: (1) on the condition that the hybridization probe and a target sequence can form a stable hybridization complex, making RNA to be detected in a substrate to contact with the hybridization probe to form the hybridization complex and (2) detecting the hybridization complex. The detection kit and the detection method provided by the invention can be used to detect the expression level of microRNA-155 gene on the level of mRNA, which is earlier than detection indexes of imaging medicine and present clinical biochemistry and can realize truly precancerous mRNA level screening. At the same time, the detection method provided by the invention is simple and convenient, requires low cost and is easy for popularization and application at country 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 mRNA 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 ...

Claims

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Application Information

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