Fluorescent cell counting detection kit for detecting tumor markers

A technology for detection kits and tumor markers, which is applied in the field of biomedical detection and can solve problems such as lack of system, mature products, and no patent authorization

Inactive Publication Date: 2018-02-09
北京金沐医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But linking it with the monitoring of solid tumors, judging from the current patent and literature search, there is still a lack of enough systematic related research, and it has not yet formed a mature product, and there is no relevant patent authorization in mainland China

Method used

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  • Fluorescent cell counting detection kit for detecting tumor markers
  • Fluorescent cell counting detection kit for detecting tumor markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] 1. Take 200ul of the peripheral blood sample of the subject to be tested, add 40ul of anti-CD14-FITC (BD company, product number 555397) and 10ul of anti-CD16-APC (BD company, product number 561304), and incubate at room temperature for 15 minutes in the dark;

[0046] 2. Add 200 ul of fixative and incubate at room temperature in the dark for 8 minutes; the components of the fixative are: formaldehyde with a volume percentage of 0.8%, and methanol with a volume percentage of 0.30%;

[0047] 3. Add 3 mL of lysate to the sample, vortex to mix, and incubate in the dark for 15 minutes to lyse red blood cells; the lyse contains 8mM Tris-HCL, 8mM NaH 2 PO 4 and / or NaHPO 4 , 140mM NaCl, 1.3% by volume TritonX-100, 8mM Na 4 P 2 o 7 10H 2 O, pH=7.2;

[0048] 4. Centrifuge at 700g for 6 minutes to remove the supernatant; pour off the supernatant slowly, absorb the residual liquid with paper, and resuspend the cells by vortexing;

[0049] 5. Add 80ul of blocking and punchin...

Embodiment 2

[0058] 1. Take 200ul of the peripheral blood sample of the subject to be tested, add 40ul of anti-CD14-FITC (BD company, product number 555397) and 10ul of anti-CD16-APC (BD company, product number 561304), and incubate at room temperature for 15 minutes in the dark;

[0059] 2. Add 200 ul of fixative and incubate at room temperature in the dark for 6 minutes; the components of the fixative are: formaldehyde with a volume percentage of 1.2%, and methanol with a volume percentage of 0.38%;

[0060] 3. Add 3 mL of lysate to the sample, vortex to mix, and incubate in the dark for 15 minutes to lyse the red blood cells; the lyse contains 12mM Tris-HCL, 12mM NaH 2 PO 4 and / or NaHPO 4 , 120mM NaCl, 0.7% by volume Triton X-100, 12mM NaCl 4 P 2 o 7 10H 2 O, pH=7.7;

[0061] 4. Centrifuge at 900g for 4 minutes to remove the supernatant; slowly pour off the supernatant, absorb the residual liquid with paper, and vortex to resuspend the cells;

[0062] 5. Add 120ul of blocking and...

Embodiment 3

[0071] 1. Take 200ul of the peripheral blood sample of the subject to be tested, add 40ul of anti-CD14-FITC (BD company, product number 555397) and 10ul of anti-CD16-APC (BD company, product number 561304), and incubate at room temperature for 15 minutes in the dark;

[0072] 2. Add 200 ul of fixative and incubate at room temperature in the dark for 5 minutes; the components of the fixative are: 1% formaldehyde by volume and 0.35% methanol by volume;

[0073] 3. Add 3 mL of lysate to the sample, vortex to mix, and incubate in the dark for 15 minutes to lyse red blood cells; the lyse contains 10mM Tris-HCL, 10mM NaH 2 PO 4 and / or NaHPO 4 , 130mM NaCl, 1% by volume TritonX-100, 10mM Na 4 P 2 o 7 10H 2 O, pH=7.5;

[0074] 4. Centrifuge at 800g for 5 minutes to remove the supernatant; pour off the supernatant slowly, absorb the residual liquid with paper, and resuspend the cells by vortexing;

[0075] 5. Add 100ul of blocking and punching buffer, and mix with fingers;

[0...

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Abstract

The present invention relates to the field of biomedical detection, particularly to a fluorescent cell counting detection kit for detecting tumor markers, wherein the fluorescent cell counting detection kit comprises a CD14 antibody and a CD16 antibody for labeling cell surface antigens, and an antibody for labeling intracytoplasm antigen, the intracytoplasm antigen is TKTL1 and/or DNASE1L1, and the kit further comprises a microscope glass slide.

Description

technical field [0001] The invention relates to the field of biomedical detection, in particular to a fluorescent cell counting detection kit for detecting tumor markers. Background technique [0002] Monocyte-macrophages include promonocytes in bone marrow, monocytes in peripheral blood, and macrophages in tissues. Human peripheral blood cells are mainly divided into anucleated cells (mainly red blood cells) and nucleated cells (white blood cells, etc.). Monocytes account for about 5% of the total number of nuclear cells and are differentiated from myeloid hematopoietic progenitor cells. They are important components of antigen-presenting cells (APC cells) in the immune system. [0003] In the peripheral blood circulation of the human body, white blood cells are distributed in two places, namely, the circulation pool and the storage pool. Circulation, but the leukocytes in the storage pool maintain a dynamic balance with the leukocytes in the circulating pool. The matura...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N21/64G01N15/14
CPCG01N33/57484G01N15/1434G01N21/6486
Inventor 郭驰张小俊杜朝阳
Owner 北京金沐医疗科技有限公司
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