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In-situ hybridization detection kit of MAGEA3 (Melanoma-Associated Antigen Antiger 3) as well as detection method and application thereof

A detection kit and in situ hybridization technology are applied in the field of kits to achieve the effects of high sensitivity, strong specificity and convenient operation

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

AI Technical Summary

Problems solved by technology

However, it has also been reported in the literature that the positive rate of MAGE-4 gene expression in liver cancer tissues of patients with HCC has nothing to do with TNM stage, tumor diameter, complete capsule, portal vein tumor thrombus, survival time, etc.

Method used

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  • In-situ hybridization detection kit of MAGEA3 (Melanoma-Associated Antigen Antiger 3) as well as detection method and application thereof
  • In-situ hybridization detection kit of MAGEA3 (Melanoma-Associated Antigen Antiger 3) as well as detection method and application thereof
  • In-situ hybridization detection kit of MAGEA3 (Melanoma-Associated Antigen Antiger 3) as well as detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] An in situ hybridization detection kit for MAGEA3 gene, comprising hybridization probes, markers, and synergists, wherein the hybridization probe sequence 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:

[0047] Digestive solution 100μl / tube 1 tube / box Colorless transparent liquid

[0048] Protective solution 100μl / tube 1 tube / box Colorless transparent liquid

[0049] Pre-hybridization solution 1300μl / tube 2 tubes / box Colorless transparent liquid

[0050] Sense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0051] Antisense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0052] Blocking solution 1000μl / tube 1 tube / box Colorless transparent liquid

[0053] Alkaline phosphatase antibody 1μl / tube 1 tube / box Colorless transparent liquid

[0054] Chromogen A 175μl / tube 1 tube / box Yellow liquid

[0055] Chromogen B...

Embodiment 2

[0097] A kind of MAGEA3 gene in situ hybridization detection method and its kit application

[0098] 1. Specimen processing

[0099] 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;

[0100] 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;

[0101] 3. Discard the supernatant. Add about twice the 1× buffer I to the precipitate, mix well, and centrifuge at 1500g / min for 10min;

[0102] 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 condi...

Embodiment 3

[0137] Parallel experiment between the detection of liver cancer with the MAGEA3 gene kit and the detection of liver cancer with the IL10 gene kit.

[0138] 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 to simultaneously detect the MAGEA3 gene and IL10 (interleukin 10) (IL10 gene sequence number NM_0005721629bp, mRNA.1q31-q32 ", cds: 60...596bp) mRNA of the gene (for nucleic acid in situ hybridization, immunohistochemical staining, microscopic counting, result reporting, etc., all adopt the in situ hybridization technology of embodiment 1 and embodiment 2 The same method and steps and reagents). It is found that the expression level of MAGEA3 gene in patients with liver cancer is higher than that of IL10 gene in patients with the same dis...

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Abstract

The invention relates to an in-situ hybridization detection kit of MAGEA3 (Melanoma-Associated Antigen Antiger 3), comprising a hybridization probe and a marker, wherein the hybridization probe sequence is shown in SEQ ID No.1. The invention also provides an in-situ hybridization detection method of the MAGEA3. In addition, the invention also provides application of the kit to the preparation of medicaments for detecting liver cancer diseases. The kit provided by the invention has the advantages of high sensitivity and strong specificity; and the detection method of the invention is convenient and simple for operation and can be widely used and popularized in municipal or higher-grade hospitals.

Description

【Technical field】 [0001] The present invention relates to a kit, in particular to an in situ hybridization detection kit for MAGEA3 gene and its detection method and application 【Background technique】 [0002] Liver cancer is one of the common malignant tumors in my country, with a high mortality rate, ranking third in the ranking of malignant tumor deaths after stomach and esophagus; in rural areas in some areas, it occupies the second place, second only to gastric cancer. About 110,000 people die from liver cancer every year in my country, accounting for 45% of the world's liver cancer deaths. The onset is often insidious, and liver cancer is often found incidentally during liver disease follow-up or physical examination. At this time, the patient is asymptomatic, and the physical examination also lacks the signs of the tumor itself. This period is called the subclinical period. Once symptoms of liver cancer appear, most of the patients who come to see a doctor have ente...

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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