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Kit and detection method for detecting CA199expression of peripheral blood circulating tumor cells of pancreatic cancer patient

A CA199 and peripheral blood circulation technology, applied in the field of kits for detecting the expression of CA199 in peripheral blood circulating tumor cells of pancreatic cancer patients, can solve problems such as bleeding, pancreatic fistula, infection, etc., to reduce cell loss, avoid edge effects, and improve accuracy Effect

Inactive Publication Date: 2020-08-21
SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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Problems solved by technology

[0005] In order to overcome the problem of multiple or real-time detection, and avoid the risks of bleeding, pancreatic fistula, and infection in biopsy, the present invention provides a kit and detection method for detecting the expression of CA199 in the peripheral blood of pancreatic cancer patients. The device is used to separate and obtain CTCs in the peripheral blood of patients with advanced or recurrent pancreatic cancer who cannot obtain tissue samples, and further use immunohistochemical technology to detect the expression of CA199 in CTCs

Method used

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  • Kit and detection method for detecting CA199expression of peripheral blood circulating tumor cells of pancreatic cancer patient
  • Kit and detection method for detecting CA199expression of peripheral blood circulating tumor cells of pancreatic cancer patient
  • Kit and detection method for detecting CA199expression of peripheral blood circulating tumor cells of pancreatic cancer patient

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Embodiment 1

[0045]1. Use a membrane filtration device to separate and obtain CTCs in the peripheral blood of patients with advanced or recurrent pancreatic cancer whose tissue samples cannot be obtained, and determine whether CTCs exist:

[0046] 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;

[0047] 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;

[0048] 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;

[0049] The diam...

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Abstract

The invention relates to a kit and a detection method for detecting the CA199 expression of peripheral blood circulating tumor cells of a pancreatic cancer patient, and belongs to the technical fieldof molecular biology. The Kit comprises 45mL of diluent, 1mL of decolorizing liquid, 0.5mL of a staining solution A, 1mL of a staining solution dyeing B, 100[mu]L of a goat anti-CA199 primary antibody, 100[mu]L of rabbit anti-goat IgG / HRP, 100[mu]L of 0.1% Triton X-100, 100[mu]L of 0.3% H2O2, 15ml of a reagent A, 50ml of a reagent B, and 60ml of a 6*PBS buffer solution. According to the detectionmethod, the CTCs in peripheral blood of a patient suffering from advanced or recurrent pancreatic cancer and incapable of obtaining a tissue specimen are separated by using a membrane filtration device, and the CA199 expression condition of the CTCs is further detected by using an immunohistochemical technology.

Description

technical field [0001] The invention relates to a kit and a detection method for detecting expression of CA199 in peripheral blood circulating tumor cells of pancreatic cancer patients, belonging to the technical field of molecular biology. Background technique [0002] Pancreatic cancer is a common malignant tumor of the digestive system, and it is also one of the malignant tumors with the worst prognosis. Among the deaths caused by malignant tumors, pancreatic cancer ranks fourth. 90% of patients die within one year after diagnosis, and the 5-year survival rate is Less than 5%. Due to the hidden and non-specific early symptoms of pancreatic cancer, strong invasiveness, early metastasis, most of them are in the middle and late stage when diagnosed, and the surgical resection rate is low, only 15% when diagnosed, so the selection of treatment methods for pancreatic cancer patients is particularly important , can make some patients avoid unnecessary surgical trauma and risk....

Claims

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Application Information

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IPC IPC(8): G01N33/574C12N5/09
CPCC12N5/0694C12N2509/00G01N33/57438G01N33/57484
Inventor 高德海崔凯李胜夏梅李浩
Owner SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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