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Kit and detection method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of gastric cancer patient

A technology of PD-L1 and peripheral blood circulation, applied in the direction of tumor/cancer cells, cell dissociation methods, animal cells, etc., can solve the problems of inability to obtain tissue samples and evaluate the status of PD-L1 of patients with tissue samples, so as to avoid the edge effect, good stability, and the effect of reducing the loss of cells

Inactive Publication Date: 2020-08-21
SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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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 puncture in real time or repeatedly, and thus to evaluate the PD-L1 status of patients, the present invention provides a non-diagnostic detection method for PD-L1 gene mutations in peripheral blood circulating tumor cells of gastric cancer patients : CTCs in the peripheral blood of patients with advanced or recurrent gastric cancer who cannot obtain tissue samples were separated by membrane filtration devices, and the expression of PD-L1 in CTCs was further detected by immunohistochemical technology

Method used

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  • Kit and detection method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of gastric cancer patient
  • Kit and detection method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of gastric cancer patient
  • Kit and detection method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of gastric cancer patient

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Embodiment 1

[0048] 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:

[0049] Collect 5ml of fasting blood for 8-12 hours from the median cubital vein, and use 45ml diluent (ingredients: 1mmol / L EDTA+0.1%BSA+0.1%trehalose+0.2%tyrosine+PBS buffer 150mmol / L, where The percentage is the volume ratio) to dilute the peripheral blood, then add 3ml of 4% paraformaldehyde to fix the diluted blood sample for 10 minutes;

[0050] 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;

[0051] Wet the filter 3 with 10ml of PBS, and then add the fixed peripheral blood sample into the blood sample container 2 of the membrane filtration device...

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Abstract

The invention discloses a kit and a detection method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of a gastric cancer patient. CTCs in peripheral blood of a patient suffering from advanced or recurrent gastric cancer and incapable of obtaining a tissue sample is obtained through separation by utilizing a membrane filtration device, and the PD-L1 expression condition of the CTCs is further detected by utilizing an immunohistochemical technology. According to the detection method provided by the invention, the PD-L1 expression condition of the patient suffering from advanced or recurrent gastric cancer can be detected without obtaining a tissue specimen through needle biopsy. The technology belongs to a minimally invasive technology and can realize real-timedetection. 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 good, the cellloss is reduced, and the detection accuracy is improved.

Description

technical field [0001] The invention provides a kit and a detection method for detecting PD-L1 gene mutations in patients with gastric cancer by circulating tumor cells in peripheral blood, and belongs 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 va...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574C12N5/09
CPCC12N5/0694C12N2509/10G01N33/57446G01N33/57484G01N2333/70532
Inventor 夏梅高德海李胜李浩
Owner SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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