In situ hybridization detection kit for early carcinoma of prostate, detecting method and use thereof

A technology for prostate cancer and detection kits, applied in the field of kits, can solve the problems of imperfect design ideas of anticancer drugs, no reports on detection technology, drug resistance of tumor cells, etc., and achieves high sensitivity, good sensitivity and specificity. strong effect

Active Publication Date: 2011-07-20
台州和和生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003]In 2005, the American Institute of Health, the Cancer Institute, the Centers for Disease Control and other units made an annual report, "Considering that human beings have failed in the war against cancer" , that is to say, the cancer mortality rate has not been reduced, and it lists several factors that lead to the failure of the anti-cancer war: 1. Tumor cell heterogeneity; 2. Tumor cell drug resistance; 3. Incomplete design of anticancer drugs, etc.
However, there is no report about the in situ hybridization detection kit and detection technology of PCADM-1 gene

Method used

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  • In situ hybridization detection kit for early carcinoma of prostate, detecting method and use thereof
  • In situ hybridization detection kit for early carcinoma of prostate, detecting method and use thereof
  • In situ hybridization detection kit for early carcinoma of prostate, detecting method and use thereof

Examples

Experimental program
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Embodiment 1

[0046] An in situ hybridization detection kit for early prostate cancer, 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:

[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...

Embodiment 2

[0097] A kind of in situ hybridization detection method of PCADM-1 gene 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 c...

Embodiment 3

[0137] A comparative experiment between the detection of prostate cancer with PCADM-1 gene kit and the detection of prostate cancer with PSA and AMACR gene kits.

[0138] In recent years, more and more prostate cancers with low expression of PSA protein have been diagnosed clinically, and the sensitivity and specificity of PSA markers in the diagnosis of prostate cancer have been increasingly questioned. PCADM-1 is a recently discovered prostate cancer-related diagnostic marker 1. PCADM-1 is only expressed in prostate cancer tissue, not in BPH, high-grade prostatic intraepithelial neoplasia and seminal vesicle tissue, and not in other tumors, and the intensity of PCADM-1 expression increases with the increase of Gleasson score (+1—— —+5). In order to scientifically evaluate the specificity, sensitivity and accuracy of PSA, AMACR gene and PCADM-1 gene in the early diagnosis of prostate cancer. We use the method of parallel experiment to detect the mRNA of PSA, AMACR and PCADM...

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Abstract

The invention relates to an in-situ hybridization detection reagent kit for early prostate cancer, which comprises a hybridization probe, a marker and a synergistic agent, wherein the hybridization probe has a sequence shown in SEQ ID NO.1. The invention also provides an in-situ hybridization detection method for a PCADM-1 gene, which comprises the following steps: a, the hybridization probe in the reagent kit contacts RNA to be detected in a substrate to form a hybridization complex; and b, the hybridization complex obtained in step a is detected. The invention also provides application of the reagent kit to preparation of a medicine for detecting early diseases of prostate cancer. The reagent kit has the advantages that the reagent kit has the characteristics of high sensitivity and strong specificity. The detection method has convenient and simple operation, and can be universally used and promoted in district hospitals and above.

Description

【Technical field】 [0001] The invention relates to a reagent kit, in particular to an in situ hybridization detection reagent kit for early prostate cancer 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 1.6 million deaths, and 6 million patients. In the world, there are 8 million new cancer patients every year, nearly 8 million deaths, and about 84 million patients. , by 2020 the number of people will double, which is a set of terrible figures. [0003] In 2005, the United States Institute of Health, Cancer Institute, Center for Disease Control and other units made an annual report, "Considering that human beings have failed in the war against cancer", that is to say, the mortality rate of cancer has not been reduced. Several factors for the failure of the cancer war are: 1. Heterogeneity of...

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 台州和和生物科技有限公司
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