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In-situ hybridization detection kit for SCCA1 genes and detection method and application thereof

A detection kit and in situ hybridization technology, applied in the field of kits, can solve problems such as unreported detection technology, and achieve the effects of high sensitivity, radical cure for malignant diseases of cancer, and convenient operation.

Inactive Publication Date: 2011-03-23
NATUREGEN BIOTECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report about the in situ hybridization detection kit and detection technology of SCCA1 gene

Method used

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  • In-situ hybridization detection kit for SCCA1 genes and detection method and application thereof
  • In-situ hybridization detection kit for SCCA1 genes and detection method and application thereof
  • In-situ hybridization detection kit for SCCA1 genes and detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] An in situ hybridization detection kit for SCCA1 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:

[0049] Digestive solution 100μl / tube 1 tube / box Colorless transparent liquid

[0050] Protective solution 100μl / tube 1 tube / box Colorless transparent liquid

[0051] Pre-hybridization solution 1300μl / tube 2 tubes / box Colorless transparent liquid

[0052] Sense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0053] Antisense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0054] Blocking solution 1000μl / tube 1 tube / box Colorless transparent liquid

[0055] Alkaline phosphatase antibody 1μl / tube 1 tube / box Colorless transparent liquid

[0056] Chromogen A 175μl / tube 1 tube / box Yellow liquid

[0057] ...

Embodiment 2

[0099] A kind of SCCA1 gene in situ hybridization detection method and its kit application

[0100] 1. Specimen processing

[0101] 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;

[0102] 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;

[0103] 3. Discard the supernatant. Add about twice the 1× buffer I to the precipitate, mix well, and centrifuge at 1500g / min for 10min;

[0104] 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

[0139] Parallel experiments between the detection of liver cancer with the SCCA1 gene kit and the detection of liver cancer with the CYK18 gene kit.

[0140] In order to scientifically evaluate the specificity, sensitivity and accuracy of the above genes in liver cancer. 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. The peripheral blood of the same patient with liver cancer was used to detect the mRNA of the SCCA1 gene and the CYK18 gene at the same time (nucleic acid in situ hybridization, immune Histochemical staining, microscopic counting, result reporting, etc. all used the same methods, steps and reagents as in the in situ hybridization technique of Example 1 and Example 2). It is found that the expression level of SCCA1 gene in patients with liver cancer is higher than that of CYK18 gene in patients with the same disease. The results showed t...

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Abstract

The invention relates to an in-situ hybridization detection kit for SCCA1 (squamous cell carcinoma antigen 1) 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 SCCA1 genes. In addition, the invention also provides application of the kit in preparation of a medicament for detecting liver cancer diseases. The kit provided by the invention has the advantages of high sensitivity and strong specificity. The detection method is convenient and simple to operate, and can be universally used and popularized in hospitals of above district level.

Description

【Technical field】 [0001] The invention relates to a reagent kit, in particular to an in situ hybridization detection reagent kit for SCCA1 gene and its detection method and application. 【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. my country is a country with a large incidence of liver cancer, accounting for 45% of the world's incidence. The high incidenc...

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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