Breast cancer early warning chip for easily rapid measuring human I type thymidine kinase gene protein

A technology for rapid determination of thymidine kinase, applied in measuring devices, biological testing, material inspection products, etc., can solve the problems of unsuitable large-scale screening, difficult application, low sensitivity, etc.

Inactive Publication Date: 2008-10-01
丁克祥
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Problems solved by technology

In short, the sensitivity of any single application of breast cancer-related tumor markers found so far is low, and the positive rates of both stages I and II are less than 30%, and early-stage patients are easy to be missed.
⑧Direct DNA sequence determination, denaturing gradient gel electrophoresis, heteroduplex analysis, fluorescent label mismatch analysis and protein truncation analysis, real-time fluorescent PCR technology, single-strand conformation polymorphism analysis, multiple heteroduplex analysis And sequencing and fluorescent double-stranded probe methods are mostly protected by patents, and because some methods are technically difficult, require high equipment, cumbersome operation steps, poor specificity, high cost, and easy pollution to cause false positives, general hospital laboratories are very difficult Difficult to apply and not suitable for large-scale screening
[0010] Even if some

Method used

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  • Breast cancer early warning chip for easily rapid measuring human I type thymidine kinase gene protein

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Experimental program
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Embodiment approach

[0052] ① Cloning and sequence analysis of hTK1 gene:

[0053] 1) Amplify TK1 cDNA by reverse transcription PCR: Extract RNA from 5 million Hela cells with Trizol kit, which can extract total ribonucleic acid (RNA) from animal cells and tissues, bacteria, fungi, etc., and treat with methotrexate 48 After purification, the precipitate was treated with a high-efficiency alkylating agent-diethyl pyrocarbonate (DEPC) to dissolve the precipitate in water to obtain 30 μl. Since DEPC can destroy ribonuclease (RNase) activity, it can remove RNase contamination.

[0054] When performing reverse transcription, add 5-fold buffer: 4 μl, primer Oligo-dT (0.5 μg / 1 μl) 1 μl, deoxynucleoside triphosphate (dNTP) at 10 mMol / L concentration: 2 μl, RNA: 10 μl, deionized water : 2 μl, mixed thoroughly, and placed in a constant temperature water bath at 70°C for 10 minutes; then, placed on ice for 2 minutes; added 1 μl reverse transcriptase (50 U, Roche), mixed well, and placed in a water bath at 4...

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Abstract

The present invention relates to a breast cancer early warning chip for measuring I type theymine deoxyriboside kinase gene protein (hTK1) easily and rapidly. The invention is characterized in that the breast cancer early warning chip is composed of the following components: a strip fiber chromatography solid phase carrier NC film which is encapsulated with an anti-hTK1 monoclone antibody (number for 2A6-McAb) that is expressed and prepared by eukaryon at one end and is encapsulated with rabbit anti-mouse IgG monoclone antibody at the other end, a filtering sample paper, a water-absorbing glass fiber paper, and a polyester diaphragm which is marked with colloidal gold and secondary antibody hTK1 monoclone antibody (number for 3E8-McAb) bonder and the like components which are adhibited on a white PVC plastic piece. The invention has the characteristics and advantages of trace quantity, high speed, sensibility, accuracy, easiness, good stability, high repeatability, strong specificity, without special equipment or device, low detecting cost, measurement to single share or a plurality shares of specimen respectively, convenient carrying and convenience for site or field general investigation or physical examination.

Description

technical field [0001] The invention relates to a breast cancer early-warning chip for the simple and rapid detection of type I thymidine kinase gene protein (hTK1), which is developed by cross-scientific technologies such as genetic engineering methods and molecular immunology techniques, and is mainly used for early warning of the risk of occurrence or development of female breast cancer At the same time, it can also be used as a new health care method for women's health protection reminder and clinical diagnosis of early breast cancer. Background technique [0002] Background on breast cancer and its diagnostic technology: Breast cancer is a common and frequently occurring malignant tumor in humans. It is a malignant tumor of epithelial origin and a systemic complex disease. The most common early clinical signs and manifestations mainly include painless lumps in the breast and armpits, changes in breast shape or size, inverted nipples, nipple discharge, "dimple-like" depr...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/532
Inventor 丁克祥刘卫国费定宇刘莉丁宇
Owner 丁克祥
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