Kit and method for detecting horizontal in situ hybridization of MICRORNA-520 at the early pathological evolution stage of a variety of cancers and application thereof

A detection kit and detection method technology, applied in the relevant detection technology field, can solve the problems of non-decreased mortality, tumor cell drug resistance, failure of the anti-cancer battle, etc., and achieve the effects of strong specificity, high sensitivity and convenient operation.

Inactive Publication Date: 2012-07-04
SUZHOU FUYING GENE TECH
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  • 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.
[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

Method used

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  • Kit and method for detecting horizontal in situ hybridization of MICRORNA-520 at the early pathological evolution stage of a variety of cancers and application thereof
  • Kit and method for detecting horizontal in situ hybridization of MICRORNA-520 at the early pathological evolution stage of a variety of cancers and application thereof
  • Kit and method for detecting horizontal in situ hybridization of MICRORNA-520 at the early pathological evolution stage of a variety of cancers and application thereof

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Experimental program
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Effect test

Embodiment 1

[0051] The in situ hybridization kit of this example was prepared according to conventional methods, and the kit included a hybridization probe designed with MICRORNA-520, a label, and instructions, wherein: the probe label of this example was digoxin.

[0052] Kit hybridization solution composition:

[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 liquid Chr...

Embodiment 2

[0061] The implementation process of applying nucleic acid in situ hybridization detection method to the expression level of MICRORNA-520 in blood samples of each group:

[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 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 a kit for detecting in situ hybridization, comprising a hybridization probe and a marker. The invention further discloses a method for detecting the in situ hybridization of MICRORNA-520 which is closely related to the early pathological evolution of a variety of cancers by using the kit. The method comprises the following steps: (1) making RNA to be detected in a substrate contact the hybridization probe to form a hybrid complex under the condition that the hybridization probe can form the stable hybrid complex together with a target sequence; and (2) detecting the hybrid complex. The kit and the method can be used for detecting the expression quantity of the MICRORNA-372 and the MICRORNA-520 on the RNA level earlier than the existing medical imaging technique and the existing clinical biochemical detection indicator and can realize the real screening of the RNA level at the earlier stage of canceration. The method is simple and convenient to operate, has low cost and can be popularized and applied to the county and district level 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 张云福裘建英裘霖张玉丽
Owner SUZHOU FUYING GENE TECH
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