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In-situ hybridization detection kit and detection method of kallikrein-related peptidase 2 (KLK2) gene and application of detection kit

A detection kit and in situ hybridization technology, applied in the field of kits, can solve the problems of lack of specificity of PSA, difficulty in distinguishing BPH and Pca, etc., and achieve the effects of radical prostate cancer, high sensitivity and convenient operation.

Inactive Publication Date: 2011-03-23
SUZHOU FUYING GENE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recently, the diagnostic accuracy of PSA has been discussed by more and more literatures. In the past, people have used prostate-specific antigen (PSA) as a marker of prostate cancer (Pca), but PSA lacks specificity. In the case of benign prostatic hyperplasia (BPH) PSA will also increase. Especially when PSA is in the range of 4-10ng / L, it is difficult to distinguish between BPH and Pca

Method used

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  • In-situ hybridization detection kit and detection method of kallikrein-related peptidase 2 (KLK2) gene and application of detection kit
  • In-situ hybridization detection kit and detection method of kallikrein-related peptidase 2 (KLK2) gene and application of detection kit
  • In-situ hybridization detection kit and detection method of kallikrein-related peptidase 2 (KLK2) gene and application of detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0045] An in situ hybridization detection kit for KLK2 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:

[0046] Digestive solution 100μl / tube 1 tube / box Colorless transparent liquid

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

[0048] Pre-hybridization solution 1300μl / tube 2 tubes / box Colorless transparent liquid

[0049] Sense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

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

[0051] Blocking solution 1000μl / tube 1 tube / box Colorless transparent liquid

[0052] Alkaline phosphatase antibody 1μl / tube 1 tube / box Colorless transparent liquid

[0053] Chromogen A 175μl / tube 1 tube / box Yellow liquid

[0054] Chro...

Embodiment 2

[0096] A KLK2 gene in situ hybridization detection method and its kit application

[0097] 1. Specimen processing

[0098] 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;

[0099] 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;

[0100] 3. Discard the supernatant. Add about twice the 1× buffer I to the precipitate, mix well, and centrifuge at 1500g / min for 10min;

[0101] 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 conditions can ...

Embodiment 3

[0136] Parallel experiment between detection of prostate cancer disease with KLK2 gene kit and detection of prostate cancer disease with ACP1 gene kit.

[0137] In order to scientifically evaluate the specificity, sensitivity and accuracy of the above genes in prostate 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, and the peripheral blood of the same patient with prostate cancer was used to simultaneously detect the KLK2 gene and ACP1 (ACP1 (acid phosphatase 1), NR_024080 ) mRNA of the gene (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 of Example 1 and Example 2). It is found that the expression level of KLK2 gene in patients with prostate cancer is higher than that of ACP1 gene in patients ...

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Abstract

The invention relates to an in-situ hybridization detection kit of a kallikrein-related peptidase 2 (KLK2 / hK2) gene. The detection kit comprises a hybridization probe and a marker, wherein the sequence of the hybridization probe is shown as SEQ ID NO.1. The invention also provides an in-situ hybridization detection method of the KLK2 gene. Moreover, the invention also provides the application of the kit to the preparation of a medicament for detecting prostatic cancer. The invention has the advantages that: the kit provided by the invention has the characteristics of high sensitivity and high specificity; and the detection method of the invention is convenient and easy to operate and can be commonly used and popularized in hospitals of district level or above.

Description

【Technical field】 [0001] The present invention relates to a kit, in particular to a KLK2 gene in situ hybridization detection kit and its detection method and application 【Background technique】 [0002] Prostate cancer (prostatic cancer) is the most common malignant tumor in the male reproductive system. The incidence increases with age, and its incidence has obvious regional differences. It is higher than lung cancer in Western Europe and North America. It is the second leading cause of cancer death in men. 1 person. The incidence rate in my country was low in the past, but due to the aging population, the incidence rate has increased in recent years, from 1.7 per 100,000 people in 1993 to 4.55 per 100,000 in 2000, and to 7.9 in 2005. Although this incidence rate is relatively low compared with other tumors, for urban men, the incidence rate of prostate cancer ranks the tenth among all tumors, which should be highly concerned. The incidence of prostate cancer is positivel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张云福裘建英
Owner SUZHOU FUYING GENE TECH
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