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mRNA (Messenger Ribose Nucleic Acid) horizontal in-situ hybridization detection kit of PITX1 at early stage of canceration and detection method and application thereof

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

Inactive Publication Date: 2012-07-11
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-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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  • mRNA (Messenger Ribose Nucleic Acid) horizontal in-situ hybridization detection kit of PITX1 at early stage of canceration and detection method and application thereof
  • mRNA (Messenger Ribose Nucleic Acid) horizontal in-situ hybridization detection kit of PITX1 at early stage of canceration and detection method and application thereof
  • mRNA (Messenger Ribose Nucleic Acid) horizontal in-situ hybridization detection kit of PITX1 at early stage of canceration and detection method and application thereof

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Effect test

Embodiment 1

[0053] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes, markers, and instructions designed with the PITX1 gene as the detection target gene, wherein:

[0054] Digoxigenin was selected as the probe label in this embodiment.

[0055] Kit hybridization solution composition:

[0056] 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...

Embodiment 2

[0064] The implementation process of applying nucleic acid in situ hybridization detection method to the expression of PITX1 gene in each group of blood samples:

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

[0066] 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;

[0067] 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;

[0068] 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 place ...

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Abstract

The invention discloses an in-situ hybridization detection kit, which comprises a hybridization probe and a marker, and discloses a method for detecting a telomerase inhibition (PITX1) gene which is closely relevant to prostatic cancer early-stage pathologic evolution, and an mRNA (Messenger Ribose Nucleic Acid) in situ in a hybridized way by using the kit. The method comprises the following steps of: (1) under the condition that a stable hybridization complex can be formed by a hybridization probe and a target sequence, contacting an RNA to be detected in a substrate with the hybridization probe to form a hybridized complex; and (2) detecting the hybridized complex. According to the kit and the detection method disclosed by the invention, the expression amount of the PITX1 gene can be detected on an mRNA level, which is earlier than image medicine and the conventional clinical biochemistry detection index, and real mRNA horizontal sieving at the early stage of canceration can be realized; and meanwhile, the detection method disclosed by the invention is easy and convenient, has low cost, and is convenient to popularize and apply in county 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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