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In situ hybridization detection kit and detection method for ICAM1 gene and application

A detection kit and in situ hybridization technology, applied in the field of kits, can solve the problems of heterogeneity, poor prognosis, drug resistance of tumor cells, etc., and achieve the effects of strong specificity, convenient operation and high sensitivity

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

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Problems solved by technology

[0002] 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 resistance of tumor cells; 3. Incomplete design of anticancer drugs, etc.
High levels of Sicam-1 are common in multiple tumors, hepatocellular carcinoma with intrahepatic dissemination, and its prognosis is poor

Method used

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  • In situ hybridization detection kit and detection method for ICAM1 gene and application
  • In situ hybridization detection kit and detection method for ICAM1 gene and application
  • In situ hybridization detection kit and detection method for ICAM1 gene and application

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Experimental program
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Effect test

Embodiment 1

[0050] An in situ hybridization detection kit for ICAM1 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:

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

[0052] Protective solution 100μl / tube 1 tube / box Colorless transparent liquid

[0053] Pre-hybridization solution 1300μl / tube 2 tubes / box Colorless transparent liquid

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

[0055] Antisense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0056] Blocking solution 1000μl / tube 1 tube / box Colorless transparent liquid

[0057] Alkaline phosphatase antibody 1μl / tube 1 tube / box Colorless transparent liquid

[0058] Chromogen A 175μl / tube 1 tube / box Yellow liquid

[0059] ...

Embodiment 2

[0101] A kind of ICAM1 gene in situ hybridization detection method and its kit application

[0102] 1. Specimen processing

[0103] 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;

[0104] 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;

[0105] 3. Discard the supernatant. Add about twice the 1× buffer I to the precipitate, mix well, and centrifuge at 1500g / min for 10min;

[0106] 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 condit...

Embodiment 3

[0142] Parallel experiment between detection of metastatic disease of esophageal cancer with ICAM1 gene kit and detection of metastatic disease of esophageal cancer with CA125 gene kit.

[0143] In order to scientifically evaluate the specificity, sensitivity and accuracy of the above genes in esophageal cancer metastasis. 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 to detect the ICAM1 gene and CA125 (CA125 (tumor marker), NM -024690) mRNA of the gene (nucleic acid in situ hybridization, immunohistochemical staining, microscopic counting, result reporting, etc. all adopt the same methods, steps and reagents as the in situ hybridization techniques of Example 1 and Example 2). It was found that the expression level of ICAM1 gene in patients with metastatic esophageal cancer was higher than that of CA125 gene in patients with the same disease. The re...

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Abstract

The invention relates to an in situ hybridization detection kit for an intracellular adhesionmolecule-1 (ICAM-1) CD-54 gene. The kit comprises a hybridization probe and a marker, wherein a sequence of the hybridization probe is shown as SEQ ID NO.1. The invention also provides an in situ hybridization detection method for the ICAM1 gene. Besides, the invention also provides application of the kit to the preparation of medicaments for detecting early metastasis of cancers and relapsed diseases. The invention has the advantages that: the kit provided by the invention has the characteristics of high sensitivity and high specificity. The detection method of the invention is convenient and easy to operate and can be widely used and popularized in hospitals of a district level or above.

Description

【Technical field】 [0001] The present invention relates to a kit, in particular to an ICAM1 gene in situ hybridization detection kit and its detection method and application 【Background technique】 [0002] 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 resistance of tumor cells; 3. Incomplete design of anticancer drugs. At the same time, the report also proposed that existing cancer diagnosis and treatment measures should be re-examined. The inventor found in the research that another two important reasons for the non-declination of the cancer mortality rate are: 1. Real early diagnosis cannot be achieved; 2. The pathological mechanism of metastasis is not clear...

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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