MRNA-level in-situ hybridization detection kit for KLOTHO gene in pancreas precancerous period of KLOTHO gene in prophase of pancreas cancer detecting method and application

A detection kit and in situ hybridization technology, which is 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 achieves convenient operation and strong specificity , the effect of high sensitivity

Inactive Publication Date: 2012-09-12
NATUREGEN BIOTECH SHANGHAI
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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-canc

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  • MRNA-level in-situ hybridization detection kit for KLOTHO gene in pancreas precancerous period of KLOTHO gene in prophase of pancreas cancer detecting method and application
  • MRNA-level in-situ hybridization detection kit for KLOTHO gene in pancreas precancerous period of KLOTHO gene in prophase of pancreas cancer detecting method and application
  • MRNA-level in-situ hybridization detection kit for KLOTHO gene in pancreas precancerous period of KLOTHO gene in prophase of pancreas cancer detecting method and application

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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 KLOTHO 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 KLOTHO gene expression of 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). 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;

[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 t...

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Abstract

The invention discloses a in-situ hybridization detection kit, comprising a hybridization probe and a marker. The invention also discloses an mRNA-level method for in-situ hybridization detection of a Klotho gene which is closely related to early pathologic evolution of pancreatic cancers by using the kit which is closely related to the pathology evolution in the prophase of pancreas cancer, comprising the following steps of, (1) under the condition that the hybridization probe and a target sequence can form a steady hybrid complex, contacting the RNA to be detected in a substrate with the hybridization probe, to form a hybrid complex; (2) detecting the hybrid complex. The kit and the detecting method of the invention can detect the expression level of the Klotho gene at mRNA level and are earlier than medical imaging and present clinic biochemistry detecting indexes. The kit and the detecting method of the invention can realize real mRNA-level screening in precancerous periods, and meanwhile, the detecting method of the invention is simple and convenient and has low cost, which makes the method easy to be applied in county 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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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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