Kit and method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of small cell lung cancer patient

A technology for small cell lung cancer and PD-L1, applied in separation methods, chemical instruments and methods, measurement devices, etc., can solve the problem of inability to assess the PD-L1 status of patients, inability to obtain tissue samples, and inability to obtain tissue samples in real time or by repeated puncture. and other problems to achieve the effect of avoiding edge effects, good stability, and reducing cell loss.

Inactive Publication Date: 2020-09-08
山东凯歌智能机器有限公司
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Problems solved by technology

[0006] In order to overcome the inability of patients with advanced or recurrent small cell lung cancer to obtain tissue samples through real-time or repeated punctures, and thus to evaluate the PD-L1 status of patients, the present invention provides a non-diagnostic method for PD-L1 gene mutations in peripheral blood circulating tumor cel

Method used

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  • Kit and method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of small cell lung cancer patient
  • Kit and method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of small cell lung cancer patient
  • Kit and method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of small cell lung cancer patient

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Embodiment 1

[0046] 1. Use a membrane filtration device to separate and obtain CTCs in the peripheral blood of patients with advanced or recurrent small cell lung cancer whose tissue samples cannot be obtained, and determine whether CTCs exist:

[0047] Collect 5ml of fasting blood for 8-12 hours from the median vein of the elbow, and use 45ml of diluent (ingredients: 1mmol / L EDTA+0.1%BSA+0.1%trehalose+0.2%polyoxyethylene polyoxypropylene ether block copolymer) Dilute the peripheral blood, then add 3ml of 4% paraformaldehyde to fix the diluted blood sample for 10 minutes;

[0048] 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;

[0049] 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 devic...

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Abstract

The invention provides a kit and method for detecting PD-L1 gene mutation of peripheral blood circulating tumor cells of a small cell lung cancer patient, and belongs to the technical field of molecular biology. The kit comprises a diluent, a decolorizing solution, a staining solution A, a staining solution B, a PD-L1 (human) primary antibody, goat anti-human IgG/HRP, 0.1% Triton X-100, 0.3% H2O2,a reagent A, a reagent B and a 6 * PBS buffer solution. According to the detection method provided by the invention, the PD-L1 expression condition of the patient suffering from advanced or recurrentsmall cell lung cancer can be detected without obtaining a tissue specimen through needle biopsy. The technology belongs to minimally invasive technology and can realize real-time detection. The false positive result caused by the edge effect possibly generated in the dyeing process can be avoided, the stability is high, the cell loss 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 mutation in peripheral blood circulating tumor cells of patients with small cell lung cancer, belonging to the technical field of molecular biology. Background technique [0002] Lung cancer is one of the main malignant tumors leading to the death of cancer patients. In my country, the morbidity and mortality of lung cancer rank first. Small cell lung cancer (SCLC) accounts for about 15% to 20% of the incidence of lung cancer. Compared with non-small cell lung cancer, it has the characteristics of faster tumor doubling time, rapid growth and early metastasis. [0003] Circulating tumor cells (CTCs) are tumor cells shed from solid tumors and enter the peripheral blood circulation. Since their discovery in 1989, a variety of methods have been used to detect circulating tumor cells in peripheral blood. Recent studies have shown that its detection has important clinical significa...

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Application Information

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IPC IPC(8): G01N33/574G01N33/569B01D29/05
CPCG01N33/57423G01N33/57492G01N33/56966B01D29/05G01N2333/70532
Inventor 侯景阳李文涛李琛琛洪瑛
Owner 山东凯歌智能机器有限公司
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