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In-situ hybridization detection kit and detection method for liver cancer precancer NOTCH1 on mRNA (messenger ribonucleic acid) level, and applications thereof

A detection kit and in situ hybridization technology, applied in the related detection technology field of mRNA expression changes, can solve problems such as non-decreasing mortality, drug resistance of tumor cells, failure of anti-cancer battle, etc., and achieve strong specificity and high sensitivity , the effect of convenient operation

Active Publication Date: 2012-07-04
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.
[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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  • In-situ hybridization detection kit and detection method for liver cancer precancer NOTCH1 on mRNA (messenger ribonucleic acid) level, and applications thereof
  • In-situ hybridization detection kit and detection method for liver cancer precancer NOTCH1 on mRNA (messenger ribonucleic acid) level, and applications thereof
  • In-situ hybridization detection kit and detection method for liver cancer precancer NOTCH1 on mRNA (messenger ribonucleic acid) level, and applications thereof

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Experimental program
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Effect test

Embodiment 1

[0054] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes, markers, and instructions designed with the NOTCH1 gene as the detection target gene, wherein:

[0055] Digoxigenin was selected as the probe label in this embodiment.

[0056] Kit hybridization solution composition:

[0057] 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 Chromoge...

Embodiment 2

[0065] The implementation process of applying the nucleic acid in situ hybridization detection method to the expression of NOTCH1 gene in each group of blood samples:

[0066] 1).Take two specimens to be tested;

[0067] 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;

[0068] 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;

[0069] 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 p...

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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 in-situ hybridization detection of mRNA (messenger ribonucleic acid) of signal conducting channel (Notch1) gene closely related to early-stage pathologic evolution of liver cancer by using the kit. The method comprises the following steps of: (1) contacting the RNA in a substrate to be detected with a hybridization probe under the condition that the hybridization probe and a target sequence can form a stable hybridization complex, so as to form the hybridization complex; and (2) detecting the hybridization complex. By adopting the kit and detection method, the expression amount of the NOTCH1 gene can be detected on the mRNA level, the detection time is earlier than that of imaging medicine and the traditional clinical biochemical detection indexes, and the mRNA-level screening and examination at the precancerous stage is truly realized; and at the same time, the detection method is simple, convenient, low in cost and convenient for popularization and application in county and district-level hospitals.

Description

technical field [0001] The present invention relates to the field of biological detection, more specifically, relates to the detection technology related to the change of mRNA expression (pathological evolution process) of precancerous changes. 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. 35% of the cost is a...

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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