Method for detecting ERCC1/CEP19 gene status based on rare cell and correlated kit

Abstract

The invention relates to a method for detecting ERCC1 / CEP19 (excision repair cross-complementing 1) gene status based on a rare cell and a correlated kit. The method comprises the following steps: step 1, acquiring a blood sample or a body fluid sample, wherein the sample contains a mixed cell population of circulating tumor cell (CTC) or other rare cells and leukocyte; step 2, using a buffer to dilute the sample, removing blood plasma, and recovering CTC or other rare cells; step 3, performing lysis by using a erythrocyte lysate, removing erythrocyte, and recovering CTC or other rare cells; step 4, uniformly mixing a magnetic particle coated with an anti-human-leucocyte correlated antigen-antibody with the recovered cell and performing incubation, so as to form a magnetic particle-leucocyte mixture through combination; step 5, centrifuging to separate the magnetic particle-leucocyte mixture from other cells, so as to obtain a mixed cell population containing CTC or other rare cells and a few of leucocytes; and step 6, using immunofluorescence cytochemistry and fluorescence in-situ hybridization combined method to determine the state of ERCC1 / CEP19 and also identify CTC and other rare cells.

Description

technical field [0001] The present invention relates to medical diagnostics, in particular to the detection of tumor cells. More specifically, the present invention relates to diagnostic methods for detecting cancer and evaluating treatment options for patients. Background technique [0002] Circulating tumor cells (Circulating Tumor Cell, CTC) refer to the tumor cells that enter the blood circulation, which can fall off into the blood from the primary tumor or metastases, and may also enter the blood before forming solid tumor lesions. Tumor cells invade the circulatory system, and most of them die within a short period of time due to the body’s immune recognition, mechanical killing, and self-apoptosis, and only a few survive and plant in distant or primary tissues and organs, and further develop into metastases. The most direct factor leading to tumor metastasis and recurrence. [0003] CTC is a "liquid biopsy" sample that can represent the primary tumor. Peripheral blo...

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Problems solved by technology

[0008] 1. As an invasive histological test limited by the location of the lesion, there are limitations such as difficulty or inability to obtain specimens, and repeated postoperative specimens cannot be obtained, resulting in the inability of tissue specimens to provide real-time monitoring information

[0009] 2. And affected by treatment, etc., the molecular information of tumor cells may change dynamically

For example, some studies have reported that breast cancer patients who received neoadjuvant chemotherapy received biopsy before chemotherapy, and compared the results with postoperative pathology, and found that the expression of ERCC1 was inconsistent before and after treatment

[0010] 3. The existence of tumor tissue heterogeneity and the fact that single-point biopsy only reflects local information makes traditional histological detection lack of comprehensive and holistic evaluation, such as comprehensive evaluation of information on primary tumors and multiple metastases, etc.

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