LET-7micro ribose nucleic acid (MIRNA) level in-situ hybridization detection kit for pathologic evolution early stage of various cancer, and detection method and application

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: 2012-07-11
NATUREGEN BIOTECH SHANGHAI
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  • 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.
[0013] 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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  • LET-7micro ribose nucleic acid (MIRNA) level in-situ hybridization detection kit for pathologic evolution early stage of various cancer, and detection method and application
  • LET-7micro ribose nucleic acid (MIRNA) level in-situ hybridization detection kit for pathologic evolution early stage of various cancer, and detection method and application
  • LET-7micro ribose nucleic acid (MIRNA) level in-situ hybridization detection kit for pathologic evolution early stage of various cancer, and detection method and application

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Embodiment 1

[0052] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes designed with LET-7MIRNA, markers, instructions, wherein:

[0053] Digoxigenin was selected as the probe label in this embodiment.

[0054] Kit hybridization solution composition:

[0055] 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...

Embodiment 2

[0063] The implementation process of applying the nucleic acid in situ hybridization detection method to the LET-7MIRNA expression level of each group of blood samples:

[0064] 1).Take two specimens to be tested;

[0065] 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;

[0066] 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;

[0067] 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 a method for performing in-situ hybridization detection on let-7micro ribose nucleic acid (miRNA) which is closely related with the pathologic evolution of the early stage of various cancer by using the kit. The method comprises the following steps of: (1) contacting RNA to be detected in substrate and the hybridization probe under the condition that the hybridization probe and a target sequence can form a stable hybridization complex to form the hybridization complex; and (2) detecting the hybridization complex. By the kit and the detection method, the let-7miRNA expression level can be detected at the RNA level; and a detection index is earlier than the detection indexes of medical imaging and the conventional clinical chemistry; real RNA-level screening of the early stage of canceration can be realized; and meanwhile, the detection method is simple and convenient, and low in cost and can be conveniently popularized and applied 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 裘霖张玉丽裘建英张云福
Owner NATUREGEN BIOTECH SHANGHAI
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