Detection kit for peripheral blood circulating tumor cells of small cell lung cancer patient and PD-L1

A technology of tumor cells and PD-LI, which is applied in the detection kit field of circulating tumor cells and PD-L1 in the peripheral blood of patients with small cell lung cancer, can solve the difficulties in evaluating the status of PD-L1, the inability to judge the status of PD-L1, and the inability to obtain tumor Tissue samples and other issues, to avoid false positive results, improve specificity, and facilitate operation

Inactive Publication Date: 2020-05-12
XIANGYA HOSPITAL CENT SOUTH UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

This method is considered to be the gold standard for PD-L1 detection, but for some patients with non-small cell lung cancer who lack surgical indications or have surgical contraindications, because the tumor tissue cannot be removed, and fine-needle puncture tissue is likely to cause tumor dissemination, PD-Ll Ll status assessment made difficult
In addition, for patients with non-small cell lung cancer metastasis and early recurrence, it is even more difficult to judge the status of PD-L1 due to the inability to obtain tumor tissue samples.

Method used

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  • Detection kit for peripheral blood circulating tumor cells of small cell lung cancer patient and PD-L1
  • Detection kit for peripheral blood circulating tumor cells of small cell lung cancer patient and PD-L1
  • Detection kit for peripheral blood circulating tumor cells of small cell lung cancer patient and PD-L1

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Experimental program
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Embodiment 1

[0059] 1. Using a microfluidic system to enrich and obtain CTCs from patients with small cell lung cancer:

[0060] 1. Acquisition of blood samples from patients with small cell lung cancer:

[0061] 1) Peripheral blood was collected from 5 patients diagnosed with small cell lung cancer, and 7.5ml of peripheral blood was collected from each case. At the same time, 7.5 ml of blood samples from 5 normal persons were taken as negative control.

[0062] 2) Peripheral blood sample pretreatment:

[0063] Use a Pasteur pipette to take 7.5ml±0.5ml of whole blood sample into the Sample Input tube, add red blood cell lysate in a corresponding proportion according to the type of blood collection tube, and centrifuge the above-mentioned lysed sample at a speed of 500g for 10 minutes. There should be precipitation at the bottom; then carefully discard the supernatant with a Pasteur pipette, retain 100 μL of the sediment and liquid at the bottom of the tube, draw 1 mL of Resuspension Buff...

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Abstract

The invention relates to a detection kit for peripheral blood circulating tumor cells of small cell lung cancer patient and PD-L1, and belongs to the technical field of peripheral blood circulating tumor cell detection reagents. A reagent for circulating tumor cell detection comprises a CTC detection primary antibody mixture and a CTC detection secondary antibody mixture, wherein the CTC detectionprimary antibody mixture comprises Anti-Cytokinase, anti-CD45, anti-pan keratin, anti-EpCAM and a Pears immunopotentiator, and the CTC detection secondary antibody mixture comprises goat anti-mouse AlexaFluor 488, goat anti-rabbit AlexaFluor 647, a DAPI working solution and a Pears immunopotentiator. The reagent provided by the invention can greatly improve the detection efficiency, reduce the detection cost and improve the detection specificity and accuracy.

Description

technical field [0001] The invention relates to the technical field of detection reagents for circulating tumor cells in peripheral blood, in particular to a detection kit for circulating tumor cells in peripheral blood of patients with small cell lung cancer and PD-L1. Background technique [0002] PD-L1 (B7-H1) belongs to the B7 family and has IgV and IgC-like regions, transmembrane regions, and cytoplasmic region tails. PD-L1 interacts with its receptor PD-1 on T cells, and plays a negative role in the immune response. It plays an important role in regulation; this molecule has a broad tissue expression profile and is highly expressed in some tumor cell lines, and many studies have shown that it is related to the immune escape mechanism of tumors. The microenvironment of the tumor site can induce the expression of PD-L1 on tumor cells, and the expression is widespread. The expressed PD-L1 is conducive to the occurrence and growth of tumors, and induces the apoptosis of an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/569
CPCG01N33/56966G01N33/57423G01N33/57492G01N2333/4742G01N2333/70525G01N2333/70532G01N2333/70589G01N2469/10
Inventor 张伟付亚成邬麟
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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