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

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 (numberfor 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 ona 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...

Examples

Embodiment approach

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

[0053] 1) Reverse transcription PCR amplification of TK1cDNA: Extract RNA from 5 million Hela cells with Trizol reagent kit that can extract total ribonucleic acid (RNA) from animal cells and tissues, bacteria, fungi, etc., and treat with methotrexate for 48 hours , and after purification, a highly efficient alkylating agent-diethyl pyrocarbonate (DEPC) was used 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 ...

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...

Examples

Embodiment approach

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

[0053] 1) Reverse transcription PCR amplification of TK1cDNA: Extract RNA from 5 million Hela cells with Trizol reagent kit that can extract total ribonucleic acid (RNA) from animal cells and tissues, bacteria, fungi, etc., and treat with methotrexate for 48 hours , and after purification, a highly efficient alkylating agent-diethyl pyrocarbonate (DEPC) was used 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 ...

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