Fluorescence immunochromatographic assay method of thymidine kinase 1 and kit

Abstract

The invention relates to a fluorescence immunochromatographic assay method of thymidine kinase 1 and a detection kit thereof. The method includes the steps of preparing a probe fixing pad, preparing an immunochromatographic test strip, preparing a sample diluting liquid and detecting a sample. The probe fixing pad is prepared by mixing thymidine kinase 1 monoclonal antibody fluorescent latex particles and goat-anti-rabbit IgG fluorescent latex particles, and diluting the mixture with a gold diluent and spraying the liquid onto glass fibers. A detection line is coated with thymidine kinase 1 monoclonal antibody and a control line is coated with goat-anti-rabbit IgG to prepare the immune-chromatographic test strip. The detection kit includes an immune-chromatographic detection card including a PVC lining plate, a sample pad, the probe fixing pad, a nitrocellulose membrane and a water absorbing paper. The probe fixing pad is prepared by mixing and drying the thymidine kinase 1 monoclonal antibody fluorescent latex particles and the goat-anti-rabbit IgG fluorescent latex particles, so that the detection kit is high in detection sensitivity and accuracy, is simple in operation and is low in cost.

Description

technical field [0001] The invention relates to a thymidine kinase 1 detection method and a kit, in particular to a thymidine kinase 1 fluorescent immunochromatographic detection method and a detection kit thereof. Background technique [0002] Thymidine kinase 1, also known as TK1, is an essential precursor for DNA synthesis in cancerous cells and a special enzyme that regulates cell synthesis, and its content is closely related to abnormal cell proliferation. Tumor is a chronic disease of abnormal cell proliferation. It is a cancer cell formed by the proliferation and abnormal differentiation of normal cells in the body under the influence of environmental factors. The continuous increase of cancer cells forms a solid tumor. However, this process is very long and often takes several years. It can only be detected by CT and other detection equipment after the time reaches a certain size. TK1 can detect these changes in the early stage of cell canceration, and then carry ou...

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Problems solved by technology

However, this method requires a long time for the immune reaction during detection. The TK1 to be tested reacts with the enzyme-labeled TK1 monoclonal antibody first, and then adds TK1-coated magnetic microspheres to undergo a competitive reaction. After washing, it is separated under a magnetic field, and finally added Fluorescent substrates are used for color development and measurement, and the steps required for detection are relatively long, and the process takes about 1.5 to 3 hours

[0006] Chinese patent CN101275954B discloses an early-warning chip that uses immunochromatography to easily measure TK1, which consists of a strip-shaped fiber chromatography solid-phase carrier NC membrane coated with an anti-TK1 monoclonal antibody, filter paper, water-absorbing glass fiber paper, and markers. The polyester film of the secondary antibody TK1 monoclonal antibody conjugate with colloidal gold is pasted on a PVC plastic sheet. This method is simple to detect, but it cannot accurately quantify TK1, and it cannot provide clear data on the monitoring of TK1 in the process of cancer treatment

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