Immunofluorescence kit and method for E-Cadherin mutation of peripheral blood circulating tumor cells of non-small cell lung cancer patient

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

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 non-small cell lung cancer to obtain tissue samples through real-time or repeated punctures, and to evaluate the status of E-Cadherin in patients, the present invention provides a method for E-Cadherin gene mutation in peripheral blood circulating tumor cells of patients w

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  • Immunofluorescence kit and method for E-Cadherin mutation of peripheral blood circulating tumor cells of non-small cell lung cancer patient
  • Immunofluorescence kit and method for E-Cadherin mutation of peripheral blood circulating tumor cells of non-small cell lung cancer patient
  • Immunofluorescence kit and method for E-Cadherin mutation of peripheral blood circulating tumor cells of non-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 non-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 d...

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Abstract

The invention provides an immunofluorescence kit and method for E-Cadherin mutation of peripheral blood circulating tumor cells of a non-small cell lung cancer patient, and belongs to the technical field of molecular biology. The kit comprises 45 mL of a diluent, 1 mL of a decolorizing solution, 0.5 mL of a staining solution A, 1 mL of a staining solution B, 200 [mu] l of methanol, 200 [mu] l of 2% PFA, 100 [mu] l of 10% goat serum, 100 [mu] l of a primary antibody working solution, 100 [mu] l of a secondary antibody working solution and a DAPI mounting medium. According to the detection method provided by the invention, the E-Cadherin expression condition of the patient suffering from advanced or recurrent non-small 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. According to the method provided by the invention, a false positive result caused by anedge 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 an immunofluorescence kit and a detection method for detecting E-Cadherin gene mutation in peripheral blood circulating tumor cells of patients with non-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. Non-small cell lung cancer (NSCLC) accounts for about 85% of all lung cancers, and more than 70% of NSCLC patients are diagnosed at an advanced stage. Despite the continuous improvement of treatment techniques such as surgery, radiotherapy and chemotherapy, the 5-year survival rate of NSCLC patients is still less than 20%, and the main causes of death include local recurrence and distant metastasis. [0003] Circulating tumor cells (CTCs) are tumor cells shed from solid tumors and enter the peripheral blood circul...

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Application Information

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IPC IPC(8): G01N33/58G01N33/574G01N33/569B01D29/05
CPCB01D29/05G01N33/56966G01N33/57423G01N33/57492G01N33/582G01N2333/705
Inventor 李琛琛洪瑛侯景阳李文涛
Owner 山东凯歌智能机器有限公司
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