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Leukemia lesion prophase mRNA level in situ hybridization detection kit, detection method and applications thereof

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

Inactive Publication Date: 2013-02-13
NATUREGEN BIOTECH SHANGHAI
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  • Summary
  • Abstract
  • 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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  • Leukemia lesion prophase mRNA level in situ hybridization detection kit, detection method and applications thereof
  • Leukemia lesion prophase mRNA level in situ hybridization detection kit, detection method and applications thereof
  • Leukemia lesion prophase mRNA level in situ hybridization detection kit, detection method and applications thereof

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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 TWIST 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 Chromogen...

Embodiment 2

[0065] The implementation process of applying the nucleic acid in situ hybridization detection method to the TWIST 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). 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 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...

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Abstract

The present invention discloses an in situ hybridization detection kit, which comprises a hybridization probe and a marker. The present invention further discloses a method for in situ hybridization detection of mRNA of transcription factor (TWIST) gene closely related with leukemia lesion prophase pathological evolution by using the kit, wherein the method comprises the following steps that: (1) RNA requiring detection in a substrate contacts a hybridization probe under a certain condition to form a hybridization complex, wherein the condition is that the hybridization probe and a target sequence can form a stable hybridization complex; and (2) the hybridization complex is detected. With the kit and the detection method, an expression level of the TWIST gene at an mRNA level can be detected so as to provide an early diagnosis compared with the existing clinical biochemical detection index, and leukemia lesion prophase mRNA level screening can be really achieved. In addition, the detection method has characteristics of simpleness, convenience and low cost, and is applicable for promotion and application in county and district hospitals.

Description

technical field [0001] The invention relates to the field of biological detection, more specifically, relates to detection technology related to the change of mRNA expression (pathological evolution process) in the pre-leukemia lesion stage. Background technique [0002] Leukemia is a malignant disease of the hematopoietic system, commonly known as "blood cancer", and is one of the top ten high-incidence malignant tumors in China. It is characterized by malignant proliferation of a certain type of leukemia cells in the hematopoietic tissue in the bone marrow or other hematopoietic tissues, and infiltrates the body Various organs and tissues lead to the inhibition of normal hematopoietic cells, resulting in various symptoms. The clinical manifestations are characterized by fever, bleeding, anemia, liver, spleen, and lymphatic enlargement. Leukemia is generally divided into acute and chronic according to the natural course of the disease and the degree of immaturity of the cel...

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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