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

A small cell lung cancer and peripheral blood circulation technology, applied in the field of molecular biology, can solve the problems of inability to obtain tissue samples, inability to evaluate patients, and inability to obtain tissue samples in real time or repeated punctures, so as to avoid edge effects, reduce cell loss, good stability effect

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 by real-time or repeated puncture, and thus to evaluate the status of E-Cadherin, the present invention provides a non-diagnostic detection method for ALK gene mutations in peripheral blood circulating tu

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

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[0046] Example 1

[0047] 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 who cannot obtain tissue specimens to determine whether CTCs exist:

[0048] Collect 5ml of fasting blood for 8-12 hours from the median cubital vein, and use 45ml of diluent (composition: 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;

[0049] In a fixed interval, 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;

[0050] Wet the filter 3 with 10ml PBS, and then add the fixed peripheral blood sample to the blood sample container 2 of the membrane filtration device to ...

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Abstract

The invention provides a kit and method for detecting E-Cadherin 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 mouse anti-human E-Cadherin monoclonal antibody, goat anti-mouse IgG/HRP, SABC, 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 E-Cadherin expression condition of the patient suffering from advanced or recurrent small cell lung cancer can be detected without obtaining a tissue specimen through needle biopsy. The technology belongs to a minimally invasive technology, and real-time detection can be realized; 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 method for detecting E-Cadherin gene mutation in peripheral blood circulating tumor cells of patients with small cell lung cancer, and belongs 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 significance...

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Application Information

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IPC IPC(8): G01N33/577G01N33/574G01N33/569B01D29/05
CPCB01D29/05G01N33/56966G01N33/57423G01N33/57492G01N33/577G01N2333/705
Inventor 洪瑛侯景阳李文涛李琛琛
Owner 山东凯歌智能机器有限公司
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