Kit and method for detecting epithelial-mesenchymal mixed circulating tumor cells

A technology for detecting kits and tumor cells, applied in the detection field of kits and epithelial-mesenchymal mixed circulating tumor cells, can solve the problems of inability to detect CTCs, limiting the clinical application effect of CTCs, and missing CTCs.

Pending Publication Date: 2019-08-20
杭州华得森生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The number of CTCs in peripheral blood is very rare, accounting for only 1/10 of peripheral blood leukocytes 6 ~1/10 7 , so the effective enrichment and detection technology bottleneck for different types of CTCs is higher and more difficult, which has always affected the practical application of CTCs
At present, the enrichment detection of CTCs mainly adopts the following two forms: ① relying on anti-EpCAM to enrich and capture CTCs and anti-cytokeratin (CK8, CK18, CK19, etc.) This method is only aimed at the enrichment and detection of epithelial CTCs, and cannot detect CTCs with mesench

Method used

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  • Kit and method for detecting epithelial-mesenchymal mixed circulating tumor cells
  • Kit and method for detecting epithelial-mesenchymal mixed circulating tumor cells
  • Kit and method for detecting epithelial-mesenchymal mixed circulating tumor cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1 (Detection kit for mixed epithelial-mesenchymal circulating tumor cells)

[0083] Such as Figure 1~3 As shown, a detection kit for epithelial-mesenchymal mixed circulating tumor cells, including a kit body 1 and a kit lid 2 matched with the kit body 1, the inner wall of the kit body 1 is closely attached to a thermal insulation and water-proof Layer 3, the bottom of the kit box body 1 is provided with a cold box 12, the cold box 12 is provided with a bottom box 4, and a plurality of placement grooves 28 and a plurality of placement holes 27 are arranged in the bottom box 4, and in the plurality of placement grooves 28 A spiral sample chamber 6 and a microfluidic chip 7 are provided, and a plurality of reagent bottles are arranged in a plurality of placement holes 27 , and the plurality of reagent bottles form a reagent bottle group 25 . The thermal insulation and water-proof layer 3 includes pearl cotton 11 , an aluminum foil 10 laminated on the pearl cotton...

Embodiment 2

[0088] Example 2 (Type detection of lung cancer circulating tumor cells and detection of PD-L1 expression)

[0089] The epithelial-mesenchymal mixed circulating tumor cell detection kit is used, and each reagent bottle in the reagent bottle group 5 is equipped with specific reagents required for detection;

[0090]Materials: The microfluidic carrier uses a microfluidic chip, and the microfluidic pump system uses a circulating tumor cell analyzer (WY-C3000, produced by Hangzhou Watson Biotechnology Co., Ltd.)

[0091] 1. Preparation of biotin-labeled epithelial-mesenchymal mixed antibody microfluidic immune capture carrier

[0092] 1.1. Use a pipette to draw 300 μL streptavidin solution with a concentration of 5 μg / mL, add it to the microfluidic chip, incubate at room temperature 25°C for 4 hours, and then place it in a drying room with an ambient humidity below 30% to dry for 24 hours until There is no liquid residue in the microfluidic chip.

[0093] 1.2. Draw 300 μL of 95%...

Embodiment 3

[0122] Example 3 CTCs Enrichment Detection Efficiency Comparison Experiment

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PUM

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Abstract

The invention discloses a kit for detecting epithelial-mesenchymic mixed circulating tumor cells, and a method for enriching and detecting epithelial-mesenchymal mixed and PD-L1 positive circulating tumor cells. The kit comprises a kit body and a kit body cover, a heat-insulating and waterproofing layer closely clings to the inner wall of the kit body, a cold box is arranged at the bottom in the kit body, a base box is arranged on the cold box, a storage grooves and a plurality of storage holes are arranged in the bottom box, spiral sample chambers and micro-fluidic chips are arranged in the storage grooves, a plurality of reagent bottles are arranged in the plurality of storage holes, and the plurality of reagent bottles form a reagent bottle group. The kit has the advantages of reasonable design combination, use and storage saving, and convenience in transportation, can be directly used for use, storage and transportation in the normal temperature environment, and ensures the effectiveness of the reagent consumable combination of the kit for detecting epithelial-mesenchymic mixed circulating tumor cells in the process for enriching, capturing and detecting epithelial, epithelial-mesenchymal mixed and mesenchyma circulating tumor cells.

Description

technical field [0001] The invention relates to the field of detection kits and epithelial-mesenchymal mixed circulating tumor cells, in particular to a detection kit for epithelial-mesenchymal mixed circulating tumor cells and enrichment detection of epithelial-mesenchymal mixed and PD-L1 positive circulating tumor cells method. Background technique [0002] Circulating Tumor Cells (CTCs) are tumor cells that are released from solid tumors or metastases into the peripheral blood circulation spontaneously or due to diagnostic and therapeutic procedures, and are a general term for various types of tumor cells that exist in peripheral blood. CTCs are mainly a heterogeneous population composed of epithelial CTCs, mixed epithelial-mesenchymal CTCs and mesenchymal CTCs subpopulations. [0003] Metastatic spread of tumors is a major cause of cancer progression and associated death. CTCs play an important role in the process of distant metastasis of tumors. If a large number of ...

Claims

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Application Information

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IPC IPC(8): B01L3/00B01L7/00B01L9/00G01N33/535G01N33/543G01N33/574
CPCB01L3/5085B01L3/50851B01L7/00B01L9/00G01N33/535G01N33/5436G01N33/574
Inventor 崔莹张开山
Owner 杭州华得森生物技术有限公司
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