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A method for detecting peripheral blood gpc3 in patients with hepatocellular carcinoma

A detection method and technology for hepatocellular carcinoma, which are applied in measurement devices, preparation and sampling of samples for testing, etc., can solve the problems that liver cancer patients are not suitable for taking tissue samples and cannot make a clear diagnosis of liver cancer, etc., and achieve the effect of real-time detection.

Active Publication Date: 2018-05-29
山东祺欣生物科技有限公司
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Problems solved by technology

[0005] In order to overcome the problem that liver cancer patients with insufficient clinical diagnosis basis are not suitable for taking tissue samples or avoid tumor dissemination caused by needle biopsy, and cannot clearly diagnose liver cancer, the present invention provides a method for detecting peripheral blood GPC3 in patients with hepatocellular carcinoma: using membrane The filter device is used to separate and obtain CTCs in the peripheral blood of patients with liver cancer who cannot obtain tissue samples, and the cell wax block technology is used to make thin-layer slices, and then the expression of GPC3 is detected

Method used

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  • A method for detecting peripheral blood gpc3 in patients with hepatocellular carcinoma
  • A method for detecting peripheral blood gpc3 in patients with hepatocellular carcinoma
  • A method for detecting peripheral blood gpc3 in patients with hepatocellular carcinoma

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Embodiment

[0052] This technique was used to isolate and identify circulating tumor cells in the peripheral blood of 8 patients with liver cancer (while detecting 8 normal samples as negative controls), and to detect the expression of GPC3.

[0053] 1. Using a membrane filtration device to separate and obtain CTCs in the peripheral blood of liver cancer patients whose tissue samples cannot be obtained:

[0054] Collect 5ml of peripheral blood from each sample, dilute the peripheral blood with 45ml diluent (ingredient: 1mmol / l EDTA+0.1%BSA), and then add 3ml of 4% paraformaldehyde to fix the diluted blood sample for 10 minutes.

[0055] At fixed intervals, assemble the membrane filtration device: as attached figure 1 , figure 2 , image 3 As shown, the filter 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 .

[0056] Wet the filter 3 with 10ml of PBS, then add the fixed peripheral blood sample into the blo...

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Abstract

The invention discloses a peripheral blood GPC3 detection method for a hepatocellular carcinoma patient. CTC in peripheral blood, where a tissue specimen can not be obtained, of the hepatocellular carcinoma patient is separated and obtained through a membrane filter device, a thin-layer slice is manufactured through the cell wax block technology, and the GPC3 expression situation is detected. By means of the technical method, the GPC3 expression situation in the peripheral blood of the hepatocellular carcinoma patient can be detected without taking hepatocellular carcinoma tissue, and the foundation is provided for hepatocellular carcinoma diagnosis. The technology is minimally invasive and even non-invasive, and real-time detection can be conducted.

Description

technical field [0001] The invention relates to a method for detecting GPC3 in peripheral blood, especially for patients with hepatocellular carcinoma who have not been pathologically diagnosed, by detecting GPC3 in peripheral blood, the diagnosis of hepatocellular carcinoma can be further confirmed. Background technique [0002] GPC3 (Glypican-3, glypican-3) is a membrane heparan sulfate proteoglycan (HSPG), with a relative molecular mass of about 66kDa, and its basic structure includes core protein, heparan sulfate (heparin sulfate, HS) chain and glycosylphosphatidylinositol (GPI), anchored on the cell surface through GPI, can bind to a variety of growth factors and regulate their activity, and then play a key role in the process of growth, development and carcinogenesis. GPC3 mainly acts through the Wnts signaling pathway, and the expression and distribution of GPC3 in normal tissues have obvious tissue specificity and phase specificity of tissue differentiation. Studies...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/78G01N1/28G01N1/36
CPCG01N1/286G01N1/36G01N21/78G01N2001/2873
Inventor 崔凯李胜王振丹
Owner 山东祺欣生物科技有限公司
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