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In-situ hybridization detection kit for MICRORNA (MICRO Ribonucleic Acid)-92A level at pathologic evolution early stage of colon cancer as well as microRNA-92A in-situ hybridization detection method and application of microRNA-92a to preparation of in-situ hybridization detection kit for colon cancer

A detection kit and in situ hybridization technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of non-decreasing mortality, drug resistance of tumor cells, failure of anti-cancer battle, etc., and achieve specificity Strong, easy to operate, high sensitivity effect

Inactive Publication Date: 2012-05-09
SUZHOU FUYING GENE TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 (MICRO Ribonucleic Acid)-92A level at pathologic evolution early stage of colon cancer as well as microRNA-92A in-situ hybridization detection method and application of microRNA-92a to preparation of in-situ hybridization detection kit for colon cancer
  • In-situ hybridization detection kit for MICRORNA (MICRO Ribonucleic Acid)-92A level at pathologic evolution early stage of colon cancer as well as microRNA-92A in-situ hybridization detection method and application of microRNA-92a to preparation of in-situ hybridization detection kit for colon cancer
  • In-situ hybridization detection kit for MICRORNA (MICRO Ribonucleic Acid)-92A level at pathologic evolution early stage of colon cancer as well as microRNA-92A in-situ hybridization detection method and application of microRNA-92a to preparation of in-situ hybridization detection kit for colon cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The in situ hybridization kit of this example was prepared according to conventional methods, and the kit included hybridization probes designed with MICRORNA-92A, markers, and instructions, wherein the probe marker 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 Chro...

Embodiment 2

[0062] The implementation process of applying nucleic acid in situ hybridization detection method to the expression level of MICRORNA-92A in each group of blood samples:

[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 a marker. The invention also discloses a method for detecting microRNA (micro Ribonucleic Acid)-92a which is closely correlated with the pathologic evolution at an early stage of colon cancer by using in-situ hybridization through the in-situ hybridization detection kit. The method comprises the following steps of: firstly, 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 to form a hybrid complex; and secondly, detecting the hybrid complex. According to the in-situ hybridization detection kit and the detection method disclosed by the invention, the expression of microRNA-92a can be detected in the RNA level, which is earlier than that by imaging medicine and traditional clinical biochemical detection index and the real RNA level screening at the early stage of canceration can be realized; and meanwhile, the detection method disclosed by the invention is simple and convenient, is low in cost and is convenience for popularization and application to county and district level 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 change of RNA expression (pathological evolution process) in the pathological evolution of colon cancer. 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 yuan....

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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