Method for specific sorting and identification detection of CTC in serum of biliary tract tumors

A specificity, tumor technology, applied in the field of biological and medical detection, to achieve the effect of improving specificity and improving sorting efficiency

Pending Publication Date: 2020-08-11
易滨 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection of CTC has expanded from simple cell counting to various directions of downstream cluster analysis, molecular typing, gene, transcription and protein level detection. It has entered clinical practice in breast cancer, colorectal cancer, prostate cancer and lung cancer. There are still a lot of gaps in the diagnosis and treatment of biliary tract cancer

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  • Method for specific sorting and identification detection of CTC in serum of biliary tract tumors
  • Method for specific sorting and identification detection of CTC in serum of biliary tract tumors

Examples

Experimental program
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Effect test

Embodiment 1

[0046] A method for specifically sorting CTCs in serum of biliary tract tumors, comprising the steps of:

[0047] (1) Sampling: Take an appropriate amount of peripheral blood from patients with biliary tract tumors, place it in an anticoagulant centrifuge tube and mix the whole blood sample;

[0048] (2) The blood sample collected in step (1) is sequentially subjected to the sample treatment of removing plasma protein and plasma nucleic acid, removing red blood cells, layered centrifugation, and removing white blood cells, specifically:

[0049] S1: Remove plasma protein and plasma nucleic acid, add the collected blood sample to the buffer, centrifuge to remove the supernatant, then gently shake the centrifuge tube to mix and precipitate the cells;

[0050] S2: Removal of red blood cells: After mixing the precipitated cells, add the lysate, place the anticoagulant centrifuge tube in a vertical mixer for mixing, discard the supernatant by centrifugation, shake the anticoagulant...

Embodiment 2

[0074] A method for specifically sorting CTCs in serum of biliary tract tumors, comprising the steps of:

[0075] (1) Sampling: Take an appropriate amount of peripheral blood from patients with biliary tract tumors, place it in an anticoagulant centrifuge tube and mix the whole blood sample;

[0076] (2) The blood sample collected in step (1) is sequentially subjected to the sample treatment of removing plasma protein and plasma nucleic acid, removing red blood cells, layered centrifugation, and removing white blood cells, specifically:

[0077] S1: Remove plasma protein and plasma nucleic acid, add the collected blood sample to the buffer, centrifuge to remove the supernatant, then gently shake the centrifuge tube to mix and precipitate the cells;

[0078] S2: Removal of red blood cells: After mixing the precipitated cells, add the lysate, place the anticoagulant centrifuge tube in a vertical mixer for mixing, discard the supernatant by centrifugation, shake the anticoagulant...

Embodiment 3

[0102] A method for specifically sorting CTCs in serum of biliary tract tumors, comprising the steps of:

[0103] (1) Sampling: Take an appropriate amount of peripheral blood from patients with biliary tract tumors, place it in an anticoagulant centrifuge tube and mix the whole blood sample;

[0104] (2) The blood sample collected in step (1) is sequentially subjected to the sample treatment of removing plasma protein and plasma nucleic acid, removing red blood cells, layered centrifugation, and removing white blood cells, specifically:

[0105] S1: Remove plasma protein and plasma nucleic acid, add the collected blood sample to the buffer, centrifuge to remove the supernatant, then gently shake the centrifuge tube to mix and precipitate the cells;

[0106] S2: Removal of red blood cells: After mixing the precipitated cells, add the lysate, place the anticoagulant centrifuge tube in a vertical mixer for mixing, discard the supernatant by centrifugation, shake the anticoagulant...

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Abstract

The present invention provides a method for specific sorting of CTC in serum of biliary tract tumors. An appropriate amount of peripheral blood from patients with biliary tract tumors is taken and placed in anticoagulation centrifuge tubes and whole blood samples are mixed evenly; the collected blood samples are sequentially subjected to sample processing of plasma protein and plasma nucleic acidremoval, red blood cell removal, layered centrifugation and white blood cell removal; an HSPG antibody or a SDC1 subtype antibody or a SDC2 subtype antibody or a GPC1 subtype antibody or a GPC3 subtype antibody is used to combine with immunomagnetic beads to capture HSPG positive or SDC1 positive or SDC2 positive or GPC1 positive or GPC3 positive CTC. HSPG is used as a relatively specific functional tumor molecular marker for CTC sorting of gallbladder cancer and cholangiocarcinoma, thereby improving sorting efficiency of biliary tract tumor CTC, a set of HSPG-based CTC molecular typing scoring system is established, the high-specificity and high-sensitivity detection method for the biliary tract tumors is otbained, and based on the high-efficiency sorting of the biliary tract tumor CTC, downstream RNA-seq and gene NGS detection is conducted to obtain tumor-specific molecular information to improve specificity.

Description

technical field [0001] The invention relates to the technical field of biological and medical detection, in particular to a method for specific sorting, identification and detection of CTC in biliary tract tumor serum. Background technique [0002] Circulating Tumor Cells (CTCs) in human peripheral blood refer to tumor cells that disseminate from tumor lesions into the peripheral blood circulation, and can develop into tumor metastatic lesions under certain conditions. Since more than 90% of cancer deaths are caused by metastasis, and CTCs are the direct source of tumor metastasis, the isolation of CTCs from blood and their molecular detection have attracted more and more attention. [0003] At present, the detection of CTC has expanded from simple cell counting to various directions of downstream cluster analysis, molecular typing, gene, transcription and protein level detection. It has entered clinical practice in breast cancer, colorectal cancer, prostate cancer and lung ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/6858
CPCC12N5/0693C12N2509/00C12Q1/6858C12Q2535/122
Inventor 易滨梁晓飞武田汪新燕刘林春
Owner 易滨
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