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Method for separating, capturing and detecting CTCs in hepatocellular carcinoma (HCC) serum

A technology of hepatocellular carcinoma and serum, which is applied in the field of biological and medical detection, can solve the problems of diagnosis and treatment limitations, and achieve the effect of improving the efficiency of sorting

Pending Publication Date: 2020-08-28
梁晓飞 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the long incubation period and extremely rapid proliferation of HCC, early diagnosis and treatment are greatly limited.

Method used

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  • Method for separating, capturing and detecting CTCs in hepatocellular carcinoma (HCC) serum
  • Method for separating, capturing and detecting CTCs in hepatocellular carcinoma (HCC) serum

Examples

Experimental program
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Effect test

Embodiment 1

[0040] A method for separating and capturing CTCs in hepatocellular carcinoma serum, comprising the steps of:

[0041] (1) Collect blood samples: take peripheral blood from patients with hepatocellular carcinoma, place it in an anticoagulant centrifuge tube and mix the whole blood sample;

[0042] (2) Pretreatment of the blood sample: the blood sample in step (1) is sequentially subjected to the processes of removing plasma protein and plasma nucleic acid, removing red blood cells, layered centrifugation, and removing white blood cells;

[0043] (3) Washing magnetic particles: pipette an appropriate amount of magnetic particle suspension combined with GPC3 antibody into the EP tube, discard the solution after standing still, add buffer and mix well, discard the supernatant after standing, repeat 3 times and use buffer Resuspend GPC3 magnetic particles to the original volume;

[0044] (4) Antibody incubation: Slowly add the magnetic particles bound to the GPC3 antibody to the ...

Embodiment 2

[0070] A method for separating and capturing CTCs in hepatocellular carcinoma serum, comprising the steps of:

[0071] (1) Collect blood samples: take peripheral blood from patients with hepatocellular carcinoma, place it in an anticoagulant centrifuge tube and mix the whole blood sample;

[0072] (2) Pretreatment of the blood sample: the blood sample in step (1) is sequentially subjected to the processes of removing plasma protein and plasma nucleic acid, removing red blood cells, layered centrifugation, and removing white blood cells;

[0073] (3) Washing magnetic particles: pipette an appropriate amount of magnetic particle suspension combined with GPC3 antibody into the EP tube, discard the solution after standing still, add buffer and mix well, discard the supernatant after standing, repeat 3 times and use buffer Resuspend GPC3 magnetic particles to the original volume;

[0074] (4) Antibody incubation: Slowly add the magnetic particles bound to the GPC3 antibody to the ...

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Abstract

The invention relates to the technical field of biological and medical detection, and provides a method for separating and capturing CTCs in hepatocellular carcinoma (HCC) serum. The method comprisesthe following steps: collecting and treating a blood sample; washing magnetic particles bound with a GPC3 antibody; antibody incubation: slowly adding the magnetic particles bound with the GPC3 antibody to the treated blood sample, adjusting the shaking speed of a shaking table, and obliquely fixing a centrifugal tube on the shaking table; and separation and capturing of the CTCs: transferring theliquid into a new anticoagulation centrifuge tube, leaning the anticoagulation centrifuge tube against a magnetic frame to enable magnetic beads for capturing the CTCs to be adsorbed on one side of the magnetic frame, inverting the magnetic frame and the anticoagulation centrifuge tube, and washing off a sample on a centrifuge tube cover. According to the method, the GPC3 antibody is bound with immunomagnetic beads to capture CTCs with positive GPC3, the sorting efficiency is improved, and the analysis method for detecting DNA in time by using a non-invasive liquid biopsy mode is realized bycombining trace evidence for learning genetic information carried by released DNA and carrying out detection and analysis.

Description

technical field [0001] The invention relates to the technical field of biological and medical detection, in particular to a method for separating, capturing and detecting CTCs in hepatocellular carcinoma serum. Background technique [0002] Hepatocellular carcinoma (Hepatocellular Carcinoma, HCC) is the main histological subtype of liver cancer, accounting for 90% of primary liver cancer, the incidence of HCC is about 10.1 cases per 100,000 people per year, and the annual death due to HCC exceeds 600,000 It is currently the sixth most common malignant tumor in the world and the second leading cause of cancer death, especially in Asia, Africa and southern Europe. Epidemiological surveys have shown that HCC in Asia is closely related to HBV infection. In addition, hepatitis C virus, liver cirrhosis, excessive alcohol consumption, and aflatoxin intake are all high-risk factors for HCC. The occurrence of HCC is a multi-stage, multi-factor synergistic effect, through the proces...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/6858
CPCC12N5/0694C12N2509/00C12Q1/6858C12Q2535/122
Inventor 梁晓飞易滨武田汪新燕刘林春
Owner 梁晓飞
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