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In-situ hybridization detection kit for MDM2 genes and detection method and application thereof

A detection kit and in situ hybridization technology, applied in the field of kits, to achieve strong specificity, high sensitivity, and the effect of radically curing breast cancer

Inactive Publication Date: 2011-03-30
NATUREGEN BIOTECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both ERK and MDM2 are well-known oncoproteins, but no interaction between them has been found so far

Method used

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  • In-situ hybridization detection kit for MDM2 genes and detection method and application thereof
  • In-situ hybridization detection kit for MDM2 genes and detection method and application thereof
  • In-situ hybridization detection kit for MDM2 genes and detection method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0044] An in situ hybridization detection kit for MDM2 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:

[0045] Digestive solution 100μl / tube 1 tube / box Colorless transparent liquid

[0046] Protective solution 100μl / tube 1 tube / box Colorless transparent liquid

[0047] Pre-hybridization solution 1300μl / tube 2 tubes / box Colorless transparent liquid

[0048] Sense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0049] Antisense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0050] Blocking solution 1000μl / tube 1 tube / box Colorless transparent liquid

[0051] Alkaline phosphatase antibody 1μl / tube 1 tube / box Colorless transparent liquid

[0052] Chromogen A 175μl / tube 1 tube / box Yellow liquid

[0053]Chr...

Embodiment 2

[0095] A kind of MDM2 gene in situ hybridization detection method and its kit application

[0096] 1. Specimen processing

[0097] 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;

[0098] 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;

[0099] 3. Discard the supernatant. Add about twice the 1× buffer I to the precipitate, mix well, and centrifuge at 1500g / min for 10min;

[0100] 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

[0135] Parallel experiments between the detection of breast cancer disease with the MDM2 gene kit and the detection of breast cancer disease with the ERK gene kit.

[0136] In order to scientifically evaluate the specificity, sensitivity and accuracy of the above genes in breast cancer. We use the method of parallel test to detect the mRNA of the above genes at the same time. The detection technology adopts nucleic acid in situ hybridization technology, and the peripheral blood of the same breast cancer patient is used to simultaneously detect the MDM2 gene and ERK (ERK oncogene, serial number NM-002745 mRNA 5916bp CDS: 241...1323bp, at chromosome 22q11.21 ".) mRNA of gene (carrying out nucleic acid in situ hybridization, immunohistochemical staining, microscopic counting, result report etc. all adopt embodiment 1 and embodiment 2 The same method and steps and reagents of in situ hybridization technique). It is found that the expression level of MDM2 gene is higher than the ex...

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Abstract

The invention relates to an in-situ hybridization detection kit for MDM2 genes, which comprises a hybridization probe and a marker, wherein the sequence of the hybridization probe is expressed as SEQ ID No.1. The invention also provides an in-situ hybridization detection method for the MDM2 genes. In addition, the invention also provides application of the kit in preparation of a medicament for detecting breast cancer diseases. The kit provided by the invention has the advantages of high sensitivity and strong specificity. The detection method is convenient and easy to operate, and can be universally used and popularized in hospitals of above district level.

Description

【Technical field】 [0001] The present invention relates to a kit, in particular to an in situ hybridization detection kit for MDM2 gene and its detection method and application 【Background technique】 [0002] Breast cancer is one of the most common malignant tumors in women. According to statistics, the incidence rate accounts for 7-10% of various malignant tumors in the whole body, and it is second only to uterine cancer in women. Its incidence is often related to heredity, and between the ages of 40 and 60, the incidence rate of women before and after menopause is higher. Only about 1-2% of breast cancer patients are men. Malignant tumors usually occurring in the glandular epithelium of the breast. It is one of the most common malignant tumors that seriously affects women's physical and mental health and even threatens their lives. [0003] It takes several years for breast cancer to change from precancer to cancer. How to achieve early detection, diagnosis, prognosis de...

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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