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Immunofluorescence kit for detecting PD-L1 gene expression of esophageal squamous cell carcinoma patient through peripheral blood circulating tumor cells

A PD-L1 and peripheral blood circulation technology, applied in the field of immunofluorescence kits, can solve the problems of false positive detection methods, tissue samples that cannot evaluate the real-time dynamic status of the patient's PD-L1 gene, false negatives, etc., to achieve good stability and avoid Edge effect, the effect of reducing the loss of cells

Inactive Publication Date: 2020-08-11
SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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Problems solved by technology

[0008] For the detection of patients with advanced or recurrent esophageal squamous cell carcinoma in the prior art, tissue samples cannot be obtained by real-time or repeated puncture, and thus the real-time dynamic status of the patient's PD-L1 gene cannot be evaluated, 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 PD-L1 gene expression of peripheral blood circulating tumor cells of patients with esophageal squamous cell carcinoma, using a membrane filtration device to separate and obtain circulating tumor cells (CTC) in the peripheral blood of patients with advanced esophageal squamous cell carcinoma , further using immunofluorescence technology to detect the expression of PD-L1 gene on CTC

Method used

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  • Immunofluorescence kit for detecting PD-L1 gene expression of esophageal squamous cell carcinoma patient through peripheral blood circulating tumor cells
  • Immunofluorescence kit for detecting PD-L1 gene expression of esophageal squamous cell carcinoma patient through peripheral blood circulating tumor cells
  • Immunofluorescence kit for detecting PD-L1 gene expression of esophageal squamous cell carcinoma patient through peripheral blood circulating tumor cells

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Embodiment 1

[0051] 1. Use a membrane filtration device to separate and obtain CTCs in the peripheral blood of patients with advanced or recurrent esophageal squamous cell carcinoma whose tissue samples cannot be obtained, and determine whether CTCs exist:

[0052] 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;

[0053] 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;

[0054] 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; ...

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Abstract

The invention provides an immunofluorescence kit for detecting PD-L1 gene expression of an esophageal squamous cell carcinoma patient through peripheral blood circulating tumor cells, and belongs to the field of molecular biology, and particularly the kit comprises a primary antibody suspension composed of goat serum, mouse anti-CK, mouse anti-CD45 and rabbit anti-PD-L1 genes, and a secondary antibody suspension composed of a fluorescently-labeled goat anti-mouse, a fluorescently-labeled goat anti-mouse and a fluorescently-labeled goat anti-rabbit. The detection method mainly comprises the following steps: collecting peripheral blood, treating the peripheral blood, filtering and enriching the circulating tumor cells, and detecting the PD-L1 gene expression condition of the circulating tumor cells by an immunofluorescence method.

Description

technical field [0001] The invention provides an immunofluorescence reagent kit for PD-L1 gene expression in peripheral blood circulating tumor cells of patients with esophageal squamous cell carcinoma, and provides a detection method, belonging to the technical field of molecular biology. Background technique [0002] Esophageal cancer is one of the most common digestive tract malignancies in the world, accounting for about 2% of all malignancies. The pathological types of esophageal cancer mainly include esophageal squamous carcinoma ESCC and esophageal adenocarcinoma EADC. More than 90% of esophageal cancer patients in my country are esophageal squamous cell carcinoma. In recent years, with the continuous improvement of diagnostic techniques for esophageal cancer, the continuous improvement of surgical operation skills, the application of preoperative neoadjuvant chemotherapy, coupled with the implementation of regular postoperative radiotherapy and chemotherapy, the qual...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574C12N5/09
CPCC12N5/0694G01N33/57407G01N33/57484G01N2333/70532
Inventor 胡敏李胜李浩于冰
Owner SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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