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Kit for AMACR gene hybridization in situ, detection method and use thereof

A detection kit and in situ hybridization technology, which is applied in the field of AMACR gene in situ hybridization detection kits, can solve the problem of unreported AMACR gene detection technology gene diagnosis kits, imperfect design ideas of anticancer drugs, and tumor cell resistance. Drug properties and other issues, to achieve the effect of high sensitivity, good sensitivity and strong specificity

Inactive Publication Date: 2009-03-18
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.
According to the existing literature, there is no report on the detection technology and gene diagnosis kit of AMACR gene

Method used

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  • Kit for AMACR gene hybridization in situ, detection method and use thereof
  • Kit for AMACR gene hybridization in situ, detection method and use thereof
  • Kit for AMACR gene hybridization in situ, detection method and use thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0049] An in situ hybridization detection kit for AMACR gene, including 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:

[0050] Digestive solution 100μl / tube 1 tube / box Colorless transparent liquid

[0051] Protective solution 100μl / tube 1 tube / box Colorless transparent liquid

[0052] Pre-hybridization solution 1300μl / tube 2 tubes / box Colorless transparent liquid

[0053] Sense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0054] Antisense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0055] Blocking solution 1000μl / tube 1 tube / box Colorless transparent liquid

[0056] Alkaline phosphatase antibody 1μl / tube 1 tube / box Colorless transparent liquid

[0057] Chromogen A 175μl / tube 1 tube / box Yellow liquid

[0058] C...

Embodiment 2

[0100] A kind of in situ hybridization detection method of AMACR gene

[0101] 1. Specimen processing

[0102] 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;

[0103] 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;

[0104] 3. Discard the supernatant. Add approximately twice the 1× buffer I to the precipitate, mix well, and centrifuge at 1500g / min for 10min;

[0105] 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 pushed to dry naturally. (Hospitals with conditions can use th...

Embodiment 3

[0139] A comparative experiment between the detection of prostate cancer with AMACR gene kit and the detection of prostate cancer with PSA and AMACR gene kit.

[0140] In recent years, more and more prostate cancers with low expression of PSA protein have been diagnosed clinically, and the sensitivity and specificity of PSA markers in the diagnosis of prostate cancer have been increasingly questioned. AMACR is a recently discovered prostate cancer-related diagnostic marker1. AMACR is only expressed in prostate cancer tissue, not in BPH, high-grade prostatic intraepithelial neoplasia and seminal vesicle tissue, and not in other tumors, and the intensity of AMACR expression increases with the increase of Gleasson score (+1—+5) . In order to scientifically evaluate the specificity, sensitivity and accuracy of PSA, AMACR gene and AMACR gene in the early diagnosis of prostate cancer. We use the method of parallel experiment to detect the mRNA of PSA, AMACR and AMACR at the same t...

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Abstract

The invention relates to a reagent kit for detecting in situ hybridization of an AMACR gene, which comprises a hybridization probe and a marker, wherein the sequence of the hybridization probe is shown as SEQ ID NO.1. The invention also provides an AMACR gene in situ hybridization detecting method. In addition, the invention also provides application of the reagent kit to the preparing and detecting of medicines for prostate gland and other cancer diseases. The reagent kit has the characteristics of high sensitivity and strong specificity. The detecting method has the advantages of simple and convenient operation, and wide application and popularization in hospitals above district level.

Description

【Technical field】 [0001] The invention relates to a kit, in particular to an in situ hybridization detection kit of AMACR gene 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. Drug resist...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 裘建英张云福
Owner NATUREGEN BIOTECH SHANGHAI
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