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AGPS gene mRNA level in-situ hybridization detection kit, detection method and applications thereof

A detection kit and in situ hybridization technology, applied in the field of biological detection, can solve problems such as human and environmental harm, and achieve the effects of convenient operation, strong specificity and high sensitivity

Inactive Publication Date: 2015-06-17
NATUREGEN BIOTECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although isotope markers have the advantages of high sensitivity and clear background, but because radioactive isotopes can cause harm to people and the environment, they have recently tended to be replaced by non-isotopes.

Method used

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  • AGPS gene mRNA level in-situ hybridization detection kit, detection method and applications thereof
  • AGPS gene mRNA level in-situ hybridization detection kit, detection method and applications thereof
  • AGPS gene mRNA level in-situ hybridization detection kit, detection method and applications thereof

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Experimental program
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Embodiment 1

[0053] Prepare the in situ hybridization kit of the present embodiment according to conventional methods, the kit includes hybridization probes designed with AGPS gene mRNA, markers, instructions, wherein:

[0054] Digoxigenin was selected as the probe label in this embodiment.

[0055] Kit hybridization solution composition:

[0056] Digestive solution 100μL / tube 1 tube / box Colorless transparent liquid

[0057] Protective solution 100μL / tube 1 tube / box Colorless transparent liquid

[0058] Pre-hybridization solution 1300μL / tube 2 tubes / box Colorless transparent liquid

[0059] Sense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0060] Antisense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0061] Blocking solution 1000μL / tube 1 tube / box Colorless transparent liquid

[0062] Alkaline phosphatase antibody 1μL / tube 1 tube / box Colorless transparent liquid

[0063] Chromogen A 175μL / tube 1 tube / box Yellow liquid

[0...

Embodiment 2

[0078] The implementation process of applying the nucleic acid in situ hybridization detection method to the AGPS gene mRNA expression of each group of blood samples:

[0079] 1).Take two specimens to be tested;

[0080] 2). Add 50 ml of digestive solution (100 μL of digestive solution plus 99.9 ml of 1× buffer Ⅰ, which is the concentration used) in a glass tank, preheat in a water bath at 37°C for 10 minutes, put 16 slides in, and treat at 37°C for 12 minutes , and then washed with 1× buffer I for 5 min;

[0081] 3). Use 0.2% protection solution (protection solution 1ml plus 1× buffer , 99ml is the concentration used), washed for 10 minutes, washed with three-distilled water for 5 minutes (the above process was carried out in a glass tank), took out the slide, and let it dry naturally;

[0082] 4). Put the slides into a humidifying box, add 25 μL / slice of pre-hybridization solution (add to the place where there are cells), cover with a cover glass, cover the humidifying bo...

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Abstract

The invention discloses an in-situ hybridization detection kit, which comprises a hybridization probe and a marker, and further discloses a method utilizing the in-situ hybridization detection kit to detect the AGPS (alkylglycerone phosphate synthase, AGPS) gene RNA expression change, which is closely related with the pathologic evolution in the early stage of canceration. The method comprises the following steps: (1) under a condition that a hybridization probe and a target sequence can form a stable hybridization complex, contacting RNA to be detected in a substrate with a hybridization probe so as to form a hybridization complex; (2) detecting the obtained hybridization complex. The provided kit and detection method can detect the expression quantity of AGPS gene in the RNA level, can detect the cancer in an earlier stage compared with the medical imaging technology and conventional clinical biochemical index detection, can achieve real RNA level screening in the early stage of canceration, moreover, have the advantages of simpleness, convenience, and low cost, and thus are convenient to promote and use in county / district hospitals.

Description

technical field [0001] The present invention relates to the field of biological detection, more specifically, relates to the detection technology related to the change of mRNA expression in the early stage of cancer pathological evolution (during pathological evolution). Background technique [0002] According to the information provided by authoritative organizations at home and abroad, the number of new cancers in my country has reached 3.12 million each year, the death toll is nearly 2.6 million, and the number of patients exceeds 7 million. There are 8 million new cancer patients every year in the world, and the death toll is close to 8 million. There are more than 84 million people, and the number will double by 2020. This is a terrible number. The cost of cancer diagnosis and treatment is getting higher and higher. The annual treatment cost of cancer patients is 200,000 yuan (may be higher in poor areas, and 200,000 yuan in developed areas). For more than 7 million p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张云福张玉丽裘建英裘霖
Owner NATUREGEN BIOTECH SHANGHAI
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