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Kit and method for detecting CEA gene mutation of peripheral blood circulating tumor cells of gastric cancer patient

A technology for peripheral blood circulation and tumor cells, applied in separation methods, chemical instruments and methods, measurement devices, etc., can solve the problems of inability to obtain tissue samples, and the inability of tissue samples to assess the CEA status of patients, avoiding edge effects and achieving good stability. , the effect of reducing the loss of cells

Inactive Publication Date: 2020-09-08
山东凯歌智能机器有限公司
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Problems solved by technology

[0006] In order to overcome the inability of patients with advanced or recurrent gastric cancer to obtain tissue samples by real-time or repeated puncture, and thus to evaluate the CEA status of patients, the present invention provides a non-diagnostic detection method for CEA gene mutations in peripheral blood circulating tumor cells of gastric cancer patients: using membrane filtration The device is used to separate and obtain CTCs in the peripheral blood of patients with advanced or recurrent gastric cancer who cannot obtain tissue samples, and further use immunohistochemical technology to detect the expression of CEA in CTCs

Method used

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  • Kit and method for detecting CEA gene mutation of peripheral blood circulating tumor cells of gastric cancer patient
  • Kit and method for detecting CEA gene mutation of peripheral blood circulating tumor cells of gastric cancer patient
  • Kit and method for detecting CEA gene mutation of peripheral blood circulating tumor cells of gastric cancer patient

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Embodiment 1

[0047] 1. Use a membrane filtration device to separate and obtain CTCs in the peripheral blood of advanced or recurrent gastric cancer patients whose tissue samples cannot be obtained, and determine whether CTCs exist:

[0048] Collect 5ml of fasting blood from the median cubital vein for 8-12 hours, and use 45ml of diluent (ingredients: 1mmol / L EDTA+0.1%BSA+0.1~10g / L soluble protein+0.1%polyglycerol fatty acid ester+PBS buffer 150mmol / L, wherein said percentage is a volume ratio) to dilute peripheral blood, then add 3ml of 4% paraformaldehyde to fix the diluted blood sample for 10 minutes;

[0049] At fixed intervals, assemble the membrane filtration device: as attached figure 1 , figure 2 , image 3 As shown, the filtering device is composed of a filter 3, a filter membrane 7, a blood sample container 2, a waste liquid tank 5, and an iron stand 1;

[0050] Wet the filter 3 with 10ml of PBS, and then add the fixed peripheral blood sample into the blood sample container 2...

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Abstract

The invention discloses a kit and method for detecting CEA gene mutation of peripheral blood circulating tumor cells of a gastric cancer patient. CTCs in peripheral blood of advanced or recurrent gastric cancer patients whose tissue specimens cannot be obtained are obtained through separation by utilizing a membrane filtration device, and CEA expression conditions of the CTCs are further detectedby utilizing an immunohistochemical technology. According to the detection method provided by the invention, the CEA expression condition of a patient suffering from advanced or recurrent gastric cancer can be detected without obtaining a tissue specimen through needle biopsy. The technology belongs to minimally invasive technology and can realize real-time detection. According to the method provided by the invention, a false positive result caused by an edge effect possibly generated in the dyeing process can be avoided, the stability is high, the cell loss is reduced, and the detection accuracy is improved.

Description

technical field [0001] The invention provides a kit and a detection method for detecting CEA gene mutation of peripheral blood circulating tumor cells of gastric cancer patients, belonging to the technical field of molecular biology. Background technique [0002] Gastric cancer (GC) is a highly fatal malignant tumor with the fourth highest incidence in the world (950,000 new cases per year), and the third highest among cancer-related deaths in 2012. Although the diagnosis and treatment methods of gastric cancer have improved in recent years, the 5-year survival rate of gastric cancer patients is less than 30%, and about 50% of gastric cancer patients suffer from tumor recurrence or metastasis after curative resection. Tumor recurrence and metastasis are the main causes of death in gastric cancer patients. [0003] Circulating tumor cells (CTCs) are tumor cells shed from solid tumors and enter the peripheral blood circulation. Since their discovery in 1989, a variety of meth...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/569B01D29/05
CPCB01D29/05G01N33/56966G01N33/57446G01N33/57473
Inventor 李琛琛洪瑛侯景阳李文涛
Owner 山东凯歌智能机器有限公司
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