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An immunofluorescence kit and detection method for detecting PD-L1 expression in peripheral blood circulating tumor cells of patients with gastric cancer

A peripheral blood circulation and PD-L1 technology, applied in the field of molecular biology, can solve the problems that tissue specimens cannot assess the real-time dynamic state of PD-L1 in patients, false positives and false negatives of detection methods, etc., to avoid edge effects and achieve good stability. , the effect of reducing the loss of cells

Active Publication Date: 2022-05-06
SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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Problems solved by technology

[0007] 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 Provides an immunofluorescence kit and detection method for the expression of PD-L1 in the peripheral blood of patients with gastric cancer, uses a membrane filtration device to separate and obtain circulating tumor cells (CTCs) in the peripheral blood of patients with advanced gastric cancer, and further uses immunofluorescence technology to detect CTCs On the expression of PD-L1

Method used

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  • An immunofluorescence kit and detection method for detecting PD-L1 expression in peripheral blood circulating tumor cells of patients with gastric cancer
  • An immunofluorescence kit and detection method for detecting PD-L1 expression in peripheral blood circulating tumor cells of patients with gastric cancer
  • An immunofluorescence kit and detection method for detecting PD-L1 expression in peripheral blood circulating tumor cells of patients with gastric cancer

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Embodiment 1

[0049] 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:

[0050] 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;

[0051] 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;

[0052] 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;

[0053] The diameter of...

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Abstract

The invention discloses an immunofluorescence kit and a detection method for detecting PD‑L1 expression of peripheral blood circulating tumor cells of patients with gastric cancer. A membrane filtration device is used to separate and obtain circulating tumor cells (CTCs) in the peripheral blood of patients with advanced gastric cancer, and further use immunofluorescence The technology detects the expression of PD-L1 on CTC. The detection method provided by the present invention can detect the expression of PD‑L1 in patients with advanced or recurrent gastric cancer without obtaining tissue samples by needle biopsy. The method provided by the invention has good separation of circulating tumor cells, can avoid the interference of blood cells, can avoid false positive results caused by possible edge effects in the staining process, has good stability, reduces cell loss, and improves detection accuracy. The use of the marker PD‑L1 showed good specificity and sensitivity, which improved its application in gastric cancer.

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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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