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In situ hybridization detection kit for PAX9 gene, and detecting method and use thereof

A detection kit and in situ hybridization technology, applied in the field of kits, can solve the problems of imperfect design ideas of anticancer drugs, unreported detection technology, drug resistance of tumor cells, etc. specific effect

Inactive Publication Date: 2009-07-01
NATUREGEN BIOTECH SHANGHAI
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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 PAX9 gene

Method used

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  • In situ hybridization detection kit for PAX9 gene, and detecting method and use thereof
  • In situ hybridization detection kit for PAX9 gene, and detecting method and use thereof
  • In situ hybridization detection kit for PAX9 gene, and detecting method and use thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0042] An in situ hybridization detection kit for PAX9 gene, comprising a hybridization probe, a marker, and a potentiator, wherein the sequence of the hybridization probe is shown in SEQ ID NO.1. Hybridization probes were labeled with digoxigenin. The composition of other liquids and specimens in the kit is as follows:

[0043] Digestive solution 100μl / tube 1 tube / box Colorless transparent liquid

[0044] Protective solution 100μl / tube 1 tube / box Colorless transparent liquid

[0045] Pre-hybridization solution 1300μl / tube 2 tubes / box Colorless transparent liquid

[0046] Sense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0047] Antisense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0048] Blocking solution 1000μl / tube 1 tube / box Colorless transparent liquid

[0049] Alkaline phosphatase antibody 1μl / tube 1 tube / box Colorless transparent liquid

[0050] Chromogen A 175μl / tube 1 tube / box Yellow liquid

[0051] C...

Embodiment 2

[0093] A kind of PAX9 gene in situ hybridization detection method and its kit application

[0094] 1. Specimen processing

[0095] 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), and centrifuge at 2000r / min 10min;

[0096] 2. Take 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;

[0097] 3. Discard the supernatant. Add about twice the 1× buffer I to the precipitate, mix well, and centrifuge at 1500g / min for 10min;

[0098] 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 conditi...

Embodiment 3

[0133] A comparative experiment between the detection of lung cancer disease with the PAX9 gene kit and the detection of lung cancer disease with the TIG2 gene kit.

[0134] In order to scientifically evaluate the specificity, sensitivity and accuracy of the above genes in lung cancer. We used the method of parallel experiments to detect the mRNA of the above genes at the same time. The detection technology used nucleic acid in situ hybridization technology. The peripheral blood of the same patient with lung cancer was used to simultaneously detect the mRNA of the PAX9 gene and the TIG2 gene (nucleic acid in situ hybridization, immune Histochemical staining, microscopic counting, result reporting, etc. all used the same methods, steps and reagents as in the in situ hybridization technique of Example 1 and Example 2). It is found that the expression level of PAX9 gene in patients with lung cancer is higher than that of TIG2 gene in patients with the same disease. The results s...

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Abstract

The invention relates to an in-situ hybridization detection reagent kit for a PAX9 gene, which comprises a hybridization probe and a marker, wherein a sequence of the hybridization probe is shown in SEQ ID NO.1. The invention also provides an in-situ hybridization detection method for the PAX9 gene. In addition, the invention also provides application of the reagent kit to preparation of a medicine for detecting a lung cancer disease. The reagent kit has the advantages that the reagent kit has the characteristics of high sensitivity and strong specificity. The detection method has convenient and simple operation, and can be universally used and promoted in district hospitals and above.

Description

【Technical field】 [0001] The invention relates to a reagent kit, in particular to a PAX9 gene in situ hybridization detection reagent kit and its detection method and application. 【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. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张云福裘建英
Owner NATUREGEN BIOTECH SHANGHAI
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