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hmg-1 gene nucleic acid in situ hybridization detection kit, detection method and application

A detection kit and in situ hybridization technology, applied in the field of cancer gene detection technology, can solve problems such as drug resistance of tumor cells, unreported HMG-1 gene detection technology, failure of the anti-cancer war, etc., to reduce mortality and the effect on morbidity

Inactive Publication Date: 2011-12-21
李学莹
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003]In 2005, the American Institute of Health, the Cancer Institute, the Centers for Disease Control and other units made an annual report, "Considering that human beings have failed in the war against cancer" , that is to say, the cancer mortality rate has not been reduced, and it lists several factors that lead to the failure of the anti-cancer war: 1. Tumor cell heterogeneity; 2. Tumor cell drug resistance; 3. Incomplete design of anticancer drugs, etc.
However, there is no report about the in situ hybridization detection kit and detection technology of HMG-1 gene

Method used

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  • hmg-1 gene nucleic acid in situ hybridization detection kit, detection method and application
  • hmg-1 gene nucleic acid in situ hybridization detection kit, detection method and application
  • hmg-1 gene nucleic acid in situ hybridization detection kit, detection method and application

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Experimental program
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Embodiment 1

[0039] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes and markers designed with the HMG-1 gene as the detection target gene, wherein:

[0040] Digoxigenin was selected as the probe label in this embodiment.

[0041] Kit composition:

[0042] Digestive solution 100μL / tube 1 tube / box Colorless transparent liquid

[0043] Protective solution 100μL / tube 1 tube / box Colorless transparent liquid

[0044] Pre-hybridization solution 1300μL / tube 2 tubes / box Colorless transparent liquid

[0045] Sense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0046] Antisense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0047] Blocking solution 1000μL / tube 1 tube / box Colorless transparent liquid

[0048] Alkaline phosphatase antibody 1μL / tube 1 tube / box Colorless transparent liquid

[0049] Chromogen A 175μL / tube 1 tube / box Yellow liquid

[0050] Chromog...

Embodiment 2

[0091] Specimen processing:

[0092] 1). Use a 10ml centrifuge tube to fill 4.5ml of lymphocyte separation solution, then slowly add 3ml of anticoagulated blood into the centrifuge tube containing lymphocyte separation solution (blood: lymphocyte separation solution = 1:1.5), 2000r / min Centrifuge for 10min

[0093] 2). Draw the white blood cells in the middle layer into another centrifuge tube, then add about twice the amount of 1× buffer I to this tube, mix well, and centrifuge at 1500g / min for 10min

[0094] 3). Discard the supernatant. Add about twice the 1× buffer I to the pellet, mix well, and centrifuge at 1500g / min for 10min

[0095] 4). Discard the supernatant, and absorb the excess liquid from the mouth of the test tube with paper towels. Then the precipitate was made into a suspension, dropped on a glass slide, and allowed to dry naturally. (Hospitals with conditions can use the film-making machine to make films.) With 3ml of blood, 4 films can be made.

[0096] ...

Embodiment 3

[0099] Prepare the reagents in the kit to the concentration used

[0100] 1). Dilute 10× buffer I with triple distilled water at 1:10 to 1× buffer I;

[0101] 2). Dilute 20× buffer II with triple distilled water at 1:10 to 2× buffer II;

[0102] Dilute 1:100 into 0.2×buffer II; dilute 1:200 into 0.1×buffer II;

[0103] 3). Dilute 10× buffer III with triple distilled water at 1:10 to 1× buffer III;

[0104] 4). Dilute 10× buffer IV with triple distilled water at a ratio of 1:10 to form × buffer IV (take 10 mL each of 1#, 2#, and 3#, and add water to 100 mL).

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Abstract

The present invention relates to an in situ hybridization detection kit for early cancer. The kit includes a hybridization probe, a label and a synergist, wherein the sequence of the hybridization probe is shown in SEQ ID NO:1. The present invention also provides an in situ hybridization detection method of HMG-1 gene, the method comprising the following steps: a. contacting the hybridization probe in the kit with the RNA to be detected in the substrate to form a hybridization complex; b. Detect the hybrid complex obtained in step a. The present invention also provides the application of the kit in the preparation of drugs for detecting early cancer diseases. The invention has the advantages that: the kit provided by the invention has the characteristics of high sensitivity and strong specificity; the detection method of the invention is convenient and simple to operate, and can be widely used and popularized in hospitals above the district level.

Description

technical field [0001] The invention relates to the field of biological detection and disease diagnosis, and more specifically relates to the gene detection technology of cancer. Background technique [0002] According to the information provided by authoritative organizations at home and abroad, there are 1.7 million new cancer cases in my country every year, nearly 1.6 million deaths, and 6 million patients. In the world, there are 8 million new cancer patients every year, nearly 8 million deaths, and about 84 million patients. , by 2020 the number of people will double, which is a set of terrible figures. [0003] In 2005, the United States Institute of Health, Cancer Institute, Center for Disease Control and other units made an annual report, "Considering that human beings have failed in the war against cancer", that is to say, the mortality rate of cancer has not been reduced. Several factors for the failure of the cancer war are: 1. Heterogeneity of tumor cells; 2. Dru...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 裘建英张云福
Owner 李学莹
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