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Detection kit for HCCR gene nucleic acid hybridization in situ, detection method and uses thereof

A detection kit and in situ hybridization technology, applied in the field of cancer gene detection technology, can solve the problem that it is difficult to confirm the only cancer focus of the tumor

Inactive Publication Date: 2011-05-25
李学莹
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This process makes it difficult to demonstrate that the mass is the only site of occurrence and a solitary focus of cancer

Method used

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  • Detection kit for HCCR gene nucleic acid hybridization in situ, detection method and uses thereof
  • Detection kit for HCCR gene nucleic acid hybridization in situ, detection method and uses thereof
  • Detection kit for HCCR gene nucleic acid hybridization in situ, detection method and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes, markers, and instructions designed with the DLK1 gene as the detection target gene, wherein:

[0037] Digoxigenin was selected as the probe label in this embodiment.

[0038] Kit composition:

[0039] Digestive solution 100μL / tube 1 tube / box Colorless transparent liquid

[0040] Protective solution 100μL / tube 1 tube / box Colorless transparent liquid

[0041] Pre-hybridization solution 1300μL / tube 2 tubes / box Colorless transparent liquid

[0042] Sense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0043] Antisense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0044]Blocking solution 1000μL / tube 1 tube / box Colorless transparent liquid

[0045] Alkaline phosphatase antibody 1μL / tube 1 tube / box Colorless transparent liquid

[0046] Chromogen A 175μL / tube 1 tube / box Yellow liquid

[...

Embodiment 2

[0088] Specimen processing:

[0089] 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

[0090] 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

[0091] 3). Discard the supernatant. Add about twice the 1× buffer I to the pellet, mix well, and centrifuge at 1500g / min for 10min

[0092] 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.

[0093] ...

Embodiment 3

[0096] Prepare the reagents in the kit to the concentration used

[0097] 1). Dilute 10× buffer I with triple distilled water at 1:10 to 1× buffer I;

[0098] 2). Dilute 20× buffer II with triple distilled water at 1:10 to 2× buffer II;

[0099] Dilute 1:100 into 0.2×buffer II; dilute 1:200 into 0.1×buffer II;

[0100] 3). Dilute 10× buffer III with triple distilled water at 1:10 to 1× buffer III;

[0101] 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 invention relates to an in situ hybridization detection kit for HCCR (Human cervical cancer oncogene binding protein) genes and early cancer. The kit comprises hybrid probes and labels, wherein the sequence of the hybrid probes is as shown by SEQ ID NO: 1. The invention also provides an in situ hybridization detection method for the HCCR genes. The detection kit and the detection method can detect the expression amount of the HCCR genes in early cancerous tumor cells or cancerous tumor cells in the genetic level, can detect change information of the HCCR genes before the protein label is generated and tumor is formed, and achieve the aim of early diagnosis of the tumor in real sense. The detection kit and the detection method have the advantages that the kit has the characteristics ofhigh sensitivity and strong specificity, and the detection method is simple and convenient to operate, and can be commonly used and popularized in hospitals with the district level and higher levels.

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 1.7 million deaths, and more than 6 million patients. Globally, there are 8 million new cancer patients every year, nearly 8 million deaths, and about 84 million patients. The number of people will more than double by 2020. The incidence and location of male cancers are arranged in order of lung, liver, and stomach; in females, they are arranged in order of breast cancer, cervical cancer, and ovarian cancer. The above cancers account for more than half of the total cancer cases. [0003] In 2005, the National Institutes of Health (NIH) Cancer Institute, Center for Disease Control and other units conducted an ann...

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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