Immunofluorescence kit for detecting PD-L1 expression of peripheral blood circulating tumor cells of gastric cancer patient and detection method

A PD-L1 and peripheral blood circulation technology, applied in tumor/cancer cells, cell dissociation methods, animal cells, etc., can solve the problem that tissue samples cannot evaluate the real-time dynamic status of PD-L1 in patients, false negatives, false positives in detection methods, etc. problems, to avoid edge effects, reduce cell loss, and achieve good stability

Active Publication Date: 2020-08-21
SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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Problems solved by technology

[0008] Aiming at the shortcomings in the prior art that the detection of patients with advanced tumors or recurrent gastric cancer cannot obtain tissue samples in real time or repeated punctures, and thus cannot evaluate the real-time dynamic state of the patient's PD-L1, and the existing detection methods are prone to false positives and false negatives, the present invention...

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  • Immunofluorescence kit for detecting PD-L1 expression of peripheral blood circulating tumor cells of gastric cancer patient and detection method
  • Immunofluorescence kit for detecting PD-L1 expression of peripheral blood circulating tumor cells of gastric cancer patient and detection method
  • Immunofluorescence kit for detecting PD-L1 expression of peripheral blood circulating tumor cells of gastric cancer patient and detection method

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Embodiment 1

[0050] 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:

[0051] Collect 5ml of fasting blood from the median vein of the elbow for 8-12 hours, dilute the peripheral blood with 45ml of diluent, and then add 3ml of 4% paraformaldehyde to fix the diluted blood sample for 10 minutes;

[0052] 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;

[0053] 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, so that it is naturally filtered by gravity, and the CTC is trapped on the filter membrane 7;

[0054] The diameter of...

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Abstract

The invention discloses an immunofluorescence kit for detecting PD-L1 expression of peripheral blood circulating tumor cells of a gastric cancer patient and a detection method. Circulating tumor cells(CTCs) in peripheral blood of an advanced gastric cancer patient are separated by using a membrane filtration device, and the PD-L1 expression condition on the CTCs is further detected by using an immunofluorescence technology. According to the detection method provided by the invention, the PD-L1 expression condition of a patient suffering from advanced or recurrent gastric cancer can be detected without obtaining a tissue specimen through needle biopsy. According to the method provided by the invention, the circulating tumor cells are well separated, the interference of blood cells can be avoided, a false positive result caused by an edge effect possibly generated in a dyeing process can be avoided, the stability is good, the loss of the cells is reduced, and the detection accuracy is improved. The marker PD-L1 shows good specificity and sensitivity in use, and the application of the marker PD-L1 in gastric cancer is improved.

Description

technical field [0001] The invention provides an immunofluorescence kit and detection method for detecting PD-L1 expression in peripheral blood circulating tumor cells of patients with gastric cancer, 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...

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Application Information

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IPC IPC(8): G01N33/574C12N5/09
CPCG01N33/57484G01N33/57446C12N5/0694G01N2333/70532C12N2509/10
Inventor 李胜李浩崔凯夏梅高德海
Owner SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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