Kit for detecting mutation expression of BRAF gene V600E of colorectal cancer patient through peripheral blood circulating tumor cells

Abstract

The invention provides a kit for detecting mutation expression of a BRAF gene V600E of a colorectal cancer patient through peripheral blood circulating tumor cells, and belongs to the field of molecular biology, and concretely the kit comprises a primary antibody suspension composed of goat serum, mouse anti-CK, mouse anti-CD45 and rabbit anti-BRAF gene V600E mutation, and a secondary antibody suspension composed of a fluorescently-labeled goat anti-mouse, a fluorescently-labeled goat anti-mouse and a fluorescently-labeled goat anti-rabbit. The detection method mainly comprises the following steps: collecting peripheral blood, treating the peripheral blood, filtering and enriching the circulating tumor cells, and detecting the mutation expression condition of the BRAF gene V600E of the circulating tumor cells by an immunofluorescence method.

Description

technical field [0001] The invention provides an immunofluorescence kit and detection method for BRAF gene V600E mutation expression of peripheral blood circulating tumor cells of colorectal cancer patients and belongs to the technical field of molecular biology. Background technique [0002] Colorectal cancer (CRC) is one of the most common digestive tract malignancies in my country. According to statistics, the morbidity and mortality of CRC are at the forefront of all malignant tumors and are on the rise. However, its pathogenic factors and pathogenesis are still not completely clear. At present, it is considered to be a complex process involving multiple genes involved in the regulation, involving the abnormal activation of oncogenes (such as RAF, etc.) and the inactivation of tumor suppressor genes (such as APC, P53, etc.). Studies have shown that BRAF gene plays an important role in the pathogenesis and treatment of colorectal cancer. However, there have not been man...

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Problems solved by technology

[0008] For the detection of patients with advanced tumors or recurrent colorectal cancer in the prior art, it is impossible to obtain tissue samples in real time or repeated punctures, and thus cannot evaluate the real-time dynamic status of the BRAF gene V600E mutation in patients, and the shortcomings of existing detection methods are prone to false positives and false negatives , the present invention provides an immunofluorescence kit and detection method for BRAF gene V600E mutation expression in peripheral blood circulating tumor cells of patients with colorectal cancer, using a membrane filtration device to separate and obtain circulating tumor cells (CTCs) in peripheral blood of patients with advanced colorectal cancer , further using immunofluorescence technology to detect the expression of BRAF gene V600E mutation on CTC

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