Method for detecting ROS1 gene status based on rare cell and correlated kit

Abstract

The invention relates to a method for detecting ROS1 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 setp 6, using immunofluorescence cytochemistry and fluorescence in-situ hybridization combined method to determine the ROS1 gene rearrangement state of CTC or other rare cells.

Description

Technical field [0001] The invention relates to medical diagnostics, in particular to the detection of tumor cells. More specifically, the present invention relates to a diagnostic method for detecting cancer and evaluating a patient's treatment plan. Background technique [0002] Circulating Tumor Cell (CTC) refers to tumor cells that enter the blood circulation, which can fall off into the blood from the primary tumor or metastasis, and may also enter the blood before the formation of solid tumor lesions. Tumor cells invade the circulatory system. Most of them die in a short period of time due to the body’s immune recognition, mechanical killing, and self-apoptosis. Only a few survive and are planted in remote or primary tissues and organs to further develop into metastases. The most direct factor leading to tumor metastasis and recurrence. [0003] As a kind of "liquid biopsy" sample that can represent the primary tumor, CTC is easy to obtain, less traumatic, and can be collec...

AI Technical Summary

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 patients with breast cancer 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 ROS1 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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