In-situ hybridization detection kit of miR-378 in early stage of pathologic evolution of gastric cancer, detection method and application

A detection kit and detection method technology, which is applied in the field of relevant detection technology for the change of RNA expression in the pathological evolution of gastric cancer, can solve the problems of human and environmental harm, and achieve the effects of convenient operation, strong specificity and high sensitivity

Inactive Publication Date: 2014-11-05
嘉兴瑞康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although isotope markers have the advantages of high sensitivity and clear background, but because radioactive isotopes can cause harm to people and the environment, they have recently tended to be replaced by non-isotopes.

Method used

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  • In-situ hybridization detection kit of miR-378 in early stage of pathologic evolution of gastric cancer, detection method and application
  • In-situ hybridization detection kit of miR-378 in early stage of pathologic evolution of gastric cancer, detection method and application
  • In-situ hybridization detection kit of miR-378 in early stage of pathologic evolution of gastric cancer, detection method and application

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Experimental program
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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 miR-378, markers, instructions, wherein:

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

[0054] Kit hybridization solution composition:

[0055]

[0056]

[0057] Concentration of prepared reagents

[0058] 1). Dilute 10× buffer I with triple distilled water 1:10 into 1× buffer I;

[0059] 2). Dilute 20× buffer II with triple distilled water 1:10 to form 2× buffer II;

[0060] Dilute 1:100 into 0.2×buffer II; dilute 1:200 into 0.1×buffer II;

[0061] 3). Dilute 10× buffer III with triple distilled water 1:10 to 1× buffer III;

[0062] 4). Dilute 10× buffer IV with triple distilled water at 1:10 to form × buffer IV (take 10 mL each of 1#, 2#, and 3#, and add water to 100 mL).

Embodiment 2

[0064] The implementation process of applying nucleic acid in situ hybridization detection method to the expression of miR-378 in blood samples of each group:

[0065] 1).Take two specimens to be tested;

[0066] 2). Add 50ml of digestive solution (100μL of digestive solution plus 1× buffer I99.9ml, 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, then Wash with 1× buffer I for 5 min;

[0067] 3). Wash with 0.2% protection solution (protection solution 1ml plus 1× buffer I, 99ml is the use concentration) for 10 minutes, three-distilled water for 5 minutes (the above process is carried out in a glass cylinder), take out the slide, let it dry naturally;

[0068] 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 box tightly, and put it in a constant temper...

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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 a method for detecting change of expression of a miR-378 gene, which is closely relative to pathologic evolution in an early stage of gastric cancer cancerization, through the kit in a manner of in-situ hybridization detection. The method includes following steps: (1) on the condition that the hybridization probe and a target sequence can form a stable hybridization complex, contacting a to-be-detected miR-378 in a substrate with the hybridization probe to form the hybridization complex; and (2) detecting the hybridization complex. By means of the kit and the method, an expression quantity of the miR-378 can be detected on a miRNA level. Compared with image medicine and a clinical biochemical detection index in the prior art, The kit and the method can be applied in an earlier stage and can achieve a true RNA-level screening operation during the early stage of cancerization. Meanwhile, the detection method is simple and convenient, is low in cost and is suitable for being popularized in hospitals in counties or districts.

Description

technical field [0001] The invention relates to the field of biological detection, more specifically, relates to detection technology related to the change of RNA expression (pathological evolution process) of gastric cancer pathological evolution. Background technique [0002] According to the information provided by authoritative organizations at home and abroad, the number of new cancers in my country has reached 3.12 million each year, the death toll is nearly 2.6 million, and the number of patients exceeds 7 million. There are 8 million new cancer patients every year in the world, and the death toll is close to 8 million. There are more than 84 million people, and the number will double by 2020. This is a terrible number. 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 i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 裘建英张云福张玉丽裘霖
Owner 嘉兴瑞康生物科技有限公司
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