DLK1 gene nucleic acid hybridization in situ detection kit, 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 problems of insufficient sensitivity of marker detection and inability to determine cancer, etc.

Inactive Publication Date: 2009-05-13
NATUREGEN BIOTECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some current biochemical diagnostic indicators are only sensitive to the detection of advanced cancers. How to make imaging medical methods unable to determine cancer, and when marker detection is not sensitive enough, we can find that cancer has occurred and has metastasized, or can predict metastasis early after treatment Recurrence situation, this is the original intention of the present invention

Method used

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  • DLK1 gene nucleic acid hybridization in situ detection kit, detection method and uses thereof
  • DLK1 gene nucleic acid hybridization in situ detection kit, detection method and uses thereof
  • DLK1 gene nucleic acid hybridization in situ detection kit, detection method and uses thereof

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Experimental program
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Effect test

Embodiment 1

[0037] 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:

[0038] Digoxigenin was selected as the probe label in this embodiment.

[0039] Kit composition:

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

[0041] Protective solution 100μL / tube 1 tube / box Colorless transparent liquid

[0042] Pre-hybridization solution 1300μL / tube 2 tubes / box Colorless transparent liquid

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

[0044] Antisense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0045] Blocking solution 1000μL / tube 1 tube / box Colorless transparent liquid

[0046] Alkaline phosphatase antibody 1μL / tube 1 tube / box Colorless transparent liquid

[0047] Chromogen A 175μL / tube 1 tube / box Yellow liquid

[...

Embodiment 2

[0089] Specimen processing:

[0090] 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

[0091] 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

[0092] 3). Discard the supernatant. Add about twice the 1× buffer I to the pellet, mix well, and centrifuge at 1500g / min for 10min

[0093] 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.

[0094] ...

Embodiment 3

[0097] Prepare the reagents in the kit to the concentration used

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

[0099] 2). Dilute 20× buffer II with triple distilled water at 1:10 to 2× buffer II;

[0100] Dilute 1:100 into 0.2×buffer II; dilute 1:200 into 0.1×buffer II;

[0101] 3). Dilute 10× buffer III with triple distilled water at 1:10 to 1× buffer III;

[0102] 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 a kit for detecting hybridization in situ for DLK1 gene of early cancer, 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 a method for detecting hybridization in situ for the DLK1 gene. The detection kit and the detection method can detect expression amount of the DLK1 gene in early canceration cells or in canceration cells at the level of genes, can detect change information of the DLK1 gene before generation of protein markers and more before oncogenesis, and really perform early diagnosis of tumor. The kit has the advantages of high sensitivity and strong specificity. The detection method has the advantage of convenient and simple operation, and can be generally used and popularized in hospitals above districts.

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 1.6 million new cancer cases in my country every year, nearly 1.6 million deaths, and 6 million patients. Globally, there are 8 million new cancer patients every year, nearly 8 million deaths, and about 84 million patients. , and the number will 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] From the onset of cancer to the survival period after treatment, taking liver cancer as an example, it is difficult to break through the five-year sur...

Claims

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Application Information

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