Method and relevant kit for detecting ALK gene status based on rare cells

Abstract

The invention relates to a method and a relevant kit for detecting ALK gene status based on rare cells. The method comprises the following steps: step 1, taking a blood sample or a body fluid sample, wherein the sample contains a mixed cell population comprising circulating tumor cells (CTCs) or other rare cells, and white blood cells; step 2, diluting the sample by adopting a buffer solution, so as to remove plasma and recover the CTCs or the other rare cells; step 3, splitting by adopting red blood cell splitting solution, so as to remove red blood cells and recover the CTCs or the other rare cells; step 4, uniformly mixing magnetic particles coated by relevant human-leukocyte-resisting antigen-antibodies and the recovered cells obtained in the step 3 for incubation, and mixing to form a magnetic particle-white blood cell mixture; step 5, separating the magnetic particle-white blood cell mixture from other cells through centrifugation, and thus obtaining the mixed cell population containing CTCs or other rare cells, and small quantity of white blood cells; and step 6, determining the ALK gene rearrangement state of CTCs or other rare cells by adopting a method of combining the immunofluorescence cytochemistry and fluorescence in situ hybridization.

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

[0011] 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

[0012] 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 there was an inconsistency in the expression of ALK before and after treatment

[0013] 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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