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Immunofluorescence kit and detection method for detecting expression of peripheral blood circulating tumor cells CA125 of gastric cancer patient

A technology of CA125 and peripheral blood circulation, applied in the field of molecular biology, can solve the problems that tissue samples cannot evaluate the real-time dynamic state of CA125 in patients, false negatives, false positives of detection methods, etc., achieve good separation of circulating tumor cells, 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

[0008] In view of the shortcomings in the prior art that the detection of patients with advanced tumors or recurrent gastric cancer cannot be obtained in real time or by repeated punctures, and the real-time dynamic state of CA125 in patients 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 expression of CA125 in the peripheral blood of gastric cancer patients, using a membrane filtration device to separate and obtain circulating tumor cells (CTCs) in the peripheral blood of patients with advanced gastric cancer, and further using immunofluorescence technology to detect the expression of CA125 on the CTCs

Method used

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  • Immunofluorescence kit and detection method for detecting expression of peripheral blood circulating tumor cells CA125 of gastric cancer patient
  • Immunofluorescence kit and detection method for detecting expression of peripheral blood circulating tumor cells CA125 of gastric cancer patient
  • Immunofluorescence kit and detection method for detecting expression of peripheral blood circulating tumor cells CA125 of gastric cancer patient

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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 and a detection method for detecting expression of peripheral blood circulating tumor cells CA125 of a gastric cancer patient. Circulating tumor cells(CTC) in peripheral blood of the patient suffering from advanced gastric cancer are obtained through separation by using a membrane filtration device, and the expression condition of CA125 on CTC isfurther detected by using an immunofluorescence technology. According to the detection method provided by the invention, the CA125 expression condition of the patient suffering from advanced or 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, 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 high, the loss of the cells is reduced, and the detection accuracy is improved. The marker CA125 shows high specificity and sensitivity in use, and application of the marker CA125 in gastric cancer is enhanced.

Description

technical field [0001] The invention provides an immunofluorescent kit and detection method for detecting expression of CA125 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 discovery ...

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Application Information

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IPC IPC(8): G01N33/58G01N33/574G01N33/569B01D29/05
CPCB01D29/05G01N33/56966G01N33/57446G01N33/57492G01N33/582G01N2333/705
Inventor 李文涛李琛琛洪瑛侯景阳
Owner 山东凯歌智能机器有限公司
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