In situ hybridization detection kit of PCNA (Proliferating Cell Nuclear Antigen) gene as well as detection method and application thereof

A detection kit and in situ hybridization technology, applied in the field of kits, can solve problems such as heterogeneity, drug resistance of tumor cells, failure of the anti-cancer war, etc., and achieve the effects of high sensitivity, strong specificity, and convenient operation

Inactive Publication Date: 2011-03-30
NATUREGEN BIOTECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] 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 tumor cells; 2. Drug resistance of tumor cells; 3. Incomplete design of anticancer drugs, etc.
The inventor found in the research that the other two important reasons for the non-declination of cancer mortality are: 1. Can not achieve real early diagnosis; 2. The pathological mechanism of metastasis is not clear

Method used

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  • In situ hybridization detection kit of PCNA (Proliferating Cell Nuclear Antigen) gene as well as detection method and application thereof
  • In situ hybridization detection kit of PCNA (Proliferating Cell Nuclear Antigen) gene as well as detection method and application thereof
  • In situ hybridization detection kit of PCNA (Proliferating Cell Nuclear Antigen) gene as well as detection method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0046] An in situ hybridization detection kit for PCNA gene, comprising a hybridization probe, a marker, and a synergist, 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

[0055] Chro...

Embodiment 2

[0097] A kind of PCNA 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 conditi...

Embodiment 3

[0138] Parallel experiment between using PCNA gene kit to detect laryngeal cancer metastases and using CA125 gene kit to detect laryngeal cancer metastases.

[0139] In order to scientifically evaluate the specificity, sensitivity and accuracy of the above genes in laryngeal cancer metastasis. 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 detect the PCNA gene and CA125 (CA125 (tumor marker), NM -024690) gene mRNA (nucleic acid in situ hybridization, immunohistochemical staining, microscopic counting, result reporting, etc. all adopt the same methods, steps and reagents as the in situ hybridization techniques in Example 1 and Example 2). It was found that the expression level of PCNA gene in patients with laryngeal cancer metastatic disease was higher than that of CA125 gene in patients with the same disease. The results showed that the specificit...

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Abstract

The invention relates to an in situ hybridization detection kit of a PCNA (Proliferating Cell Nuclear Antigen) gene, comprising a hybridization probe and a marker, wherein the sequence of the hybridization probe is shown as SEQID NO.1. The invention also provides an in situ hybridization detection method of the PCNA gene and additionally provides application of the kit in preparing a drug for detecting cancer early metastasis and recrudescent diseases. The kit has the advantages of high sensitivity and strong specificity. The detection method is convenient and simple for operation and can be universally used and popularized in hospitals above a district level.

Description

【Technical field】 [0001] The present invention relates to a kit, in particular to a PCNA gene in situ hybridization detection kit and its detection method and application 【Background technique】 [0002] 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 tumor cells; 2. Drug resistance of tumor cells; 3. Incomplete design of anticancer drugs. At the same time, the report also proposed that existing cancer diagnosis and treatment measures should be re-examined. The inventor found in the research that another two important reasons for the non-declination of the cancer mortality rate are: 1. Real early diagnosis cannot be achieved; 2. The pathological mechanism of metastasis is not clear. ...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12Q1/68
Inventor裘建英张云福
OwnerNATUREGEN BIOTECH SHANGHAI