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Kit and detection method for detecting PD-L1 expression of peripheral blood circulating tumor cells of pancreatic cancer patient

A technology of PD-L1 and peripheral blood circulation, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as pancreatic fistula and infection, reduce cell loss, avoid edge effects, and improve accuracy

Inactive Publication Date: 2020-08-18
SHANDONG RES INST OF TUMOUR PREVENTION TREATMENT
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In order to solve the problem of real-time dynamic detection of PD-L1 expression, and avoid the risks of bleeding, pancreatic fistula, and infection in biopsy, the present invention provides a non-diagnostic detection method for PD-L1 expression in peripheral blood circulating tumor cells of pancreatic cancer patients. CTCs in the peripheral blood of patients with advanced or recurrent pancreatic cancer who could not 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 expression of peripheral blood circulating tumor cells of pancreatic cancer patient
  • Kit and detection method for detecting PD-L1 expression of peripheral blood circulating tumor cells of pancreatic cancer patient
  • Kit and detection method for detecting PD-L1 expression of peripheral blood circulating tumor cells of pancreatic 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 patients with advanced or recurrent pancreatic cancer whose tissue samples cannot be obtained, and determine whether CTCs exist:

[0049] 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;

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

[0052] The dia...

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Abstract

The invention relates to a kit and a detection method for detecting PD-L1 expression of peripheral blood circulating tumor cells of a pancreatic cancer patient, and belongs to the technical field of molecular biology. The kit comprises 45mL of a diluent, 1mL of decolorizing liquid, 0.5 mL of staining solution A, 1mL of dyeing liquid B, 100 [mu]L of an anti-human PD-L1 primary antibody, 100 [mu]L of goat anti-human IgG / HRP, 100 [mu]L of 0.1% Triton X-100, 100 [mu]L of 0.3% H2O2, 15 ml of a reagent A, 50 ml of a reagent B, and 60 mL of a 6*PBS buffer solution. CTCs in peripheral blood of advanced or recurrent pancreatic cancer patients who cannot obtain tissue specimens are 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.

Description

technical field [0001] The invention relates to a kit and a detection method for detecting the expression of PD-L1 in peripheral blood circulating tumor cells of pancreatic cancer patients, belonging to the technical field of molecular biology. Background technique [0002] Pancreatic cancer is a common malignant tumor of the digestive system, and it is also one of the malignant tumors with the worst prognosis. Among the deaths caused by malignant tumors, pancreatic cancer ranks fourth. 90% of patients die within one year after diagnosis, and the 5-year survival rate is Less than 10%. Due to the hidden and non-specific early symptoms of pancreatic cancer, strong invasiveness, early metastasis, and low surgical resection rate, the disease is mostly in the middle and late stages when it is diagnosed. Currently, systemic chemotherapy is basically the treatment for pancreatic cancer. [0003] Circulating tumor cells (CTCs) are tumor cells shed from solid tumors and enter the pe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/57438G01N33/57492G01N2333/70532
Inventor 崔凯李胜黄宁胡敏戚元刚
Owner SHANDONG RES INST OF TUMOUR PREVENTION TREATMENT
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