Immunofluorescence kit for E-Cadherin expression of peripheral blood circulating tumor cells of gastric cancer patient and detection method
A technology for peripheral blood circulation and tumor cells, applied in separation methods, chemical instruments and methods, biological tests, etc., can solve the problems of false negatives, the inability of tissue specimens to evaluate the real-time dynamic status of E-Cadherin in patients, and false positives of detection methods. Reduce cell loss, avoid edge effects, good stability
Inactive Publication Date: 2020-09-11
山东凯歌智能机器有限公司
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[0008] Aiming at the shortcomings in the prior art that it is impossible to obtain tissue samples by real-time or repeated punctures for the detection of patients with advanced tumors or recurrent gastric cancer, and thus cannot evaluate the real-time dynamic state of E-Cadherin in patients, 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 expression of E-Cadherin in the peripheral blood of patients with gastric cancer, uses a membrane filtration device to separate and obtain circulating tumor cells (CTCs) in the peripheral blood of patients with advanced gastric cancer, and further uses immunofluorescence technology to detect CTCs Expression of E-Cadherin
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Embodiment 1
[0050] 1. Use a membrane filtration device to separate and obtain CTCs in the peripheral blood of advanced or recurrent gastric cancer patients whose tissue samples cannot be obtained, and determine whether CTCs exist:
[0051] 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;
[0052] 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;
[0053] 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;
[0054] The diameter of...
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Abstract
The invention discloses an immunofluorescence kit for detecting E-Cadherin expression of peripheral blood circulating tumor cells of a gastric cancer patient and a detection method. Circulating tumorcells (CTC) in peripheral blood of a patient suffering from advanced gastric cancer are obtained through separation by utilizing a membrane filtration device, and the expression condition of E-Cadherin on CTC is further detected by utilizing an immunofluorescence technology. According to the detection method provided by the invention, the E-Cadherin expression condition of a patient with advancedor recurrent gastric cancer can be detected without obtaining a tissue specimen through needle biopsy. According to the method provided by the invention, the circulating tumor cells are well separated, the interference of blood cells can be avoided, a false positive result caused by an edge effect possibly generated in a dyeing process can be avoided, the stability is good, the loss of the cells is reduced, and the detection accuracy is improved. The marker E-Cadherin shows good specificity and sensitivity in use, and the application of the marker E-Cadherin in gastric cancer is improved.
Description
technical field [0001] The invention provides an immunofluorescence kit and detection method for detecting the expression of E-Cadherin in peripheral blood circulating tumor cells of patients with gastric cancer, and belongs to the technical field of molecular biology. Background technique [0002] Gastric cancer (GC) is a highly fatal malignant tumor with the fourth highest incidence in the world (950,000 new cases per year), and the third highest among cancer-related deaths in 2012. Although the diagnosis and treatment methods of gastric cancer have improved in recent years, the 5-year survival rate of gastric cancer patients is less than 30%, and about 50% of gastric cancer patients suffer from tumor recurrence or metastasis after curative resection. Tumor recurrence and metastasis are the main causes of death in gastric cancer patients. [0003] Circulating tumor cells (CTCs) are tumor cells shed from solid tumors and enter the peripheral blood circulation. Since their ...
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IPC IPC(8): G01N33/58G01N33/574G01N33/569B01D29/05
CPCB01D29/05G01N33/56966G01N33/57446G01N33/57492G01N33/582G01N2333/705
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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