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Bladder cancer detection device and method

A technology for bladder cancer cells and cancer cells, which is applied in the field of bladder cancer detection devices and methods, and can solve the problems of increasing sample analysis time and cost, reducing background, etc.

Inactive Publication Date: 2020-01-31
UNIVERSITY OF SOUTH AUSTRALIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(OliveiraArcolino 50 et al 2015) In order to minimize background, natural urine has to be processed extensively, a process that increases sample analysis time and cost

Method used

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  • Bladder cancer detection device and method
  • Bladder cancer detection device and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-96

[0107] Example 1- Preparation of a 96-well plate plasma polymer substrate

[0108] For cell capture, 96-well plates (Corning, Costar) and microscope grade glass slides (Objekttrager) are used as solid substrates. Use silicon wafers for plasma polymer film thickness measurement.

[0109] Before plasma deposition, the glass slides and silicon wafers were washed in piranha solution (H2O2:H2SO4, 1:3), and rinsed thoroughly with milliQ ultrapure water. Use a sterile 96-well plate as obtained.

[0110] The oxazoline-based thin film coating is deposited on the solid substrate by continuous plasma deposition as described previously. (Macgregor-Ramiasa et al. 2015b; Ramiasa et al. 2015) Simply put, the customized parallel plate plasma reactor is placed in a vacuum (2.10 -2 mbar), and inoculate 2-methyl-2-oxazoline precursor (Sigma-Aldrich, Australia) with a needle valve in the chamber until a constant monomer flow rate of 8 standard cubic centimeters per minute (sccm) is reached. Then the ...

Embodiment 2

[0113] Example 2-Preparation of Microfluidic Plasma Polymer Substrate

[0114] The method described in Example 1 was used to form a PPOx-coated microscope slide. Then use standard techniques on the PPOx coated microscope slide and Ibidi IV 0.4 sticky (DSKH) Micro-channels are formed between the sticky-tear glass slides.

Embodiment 3

[0115] Example 3-Covalent attachment of antibody to plasma polymer substrate

[0116] The cell capture "chamber" consists of individual wells (Example 1) or microchannels (Example 2) in a standard 96-well plate. The unique reactivity of plasma-deposited polyoxazoline and carboxylic acid is used to irreversibly bind antibodies to the plasma polymer substrate. (Macgregor-Ramiasa et al. 2015a; Schmidt et al. 1994; Tillet et al. 2011) Dissolve Anti-EpCAM antibody in 100% PBS at 10 μg.mL-1, gently pipette 50 μL onto the plasma polymer substrate and make They were combined overnight at 4°C. Then draw the antibody solution and pass in 0.1 mg.mL -1 The surface of the active polymer was blocked with skim milk by incubating for 15 minutes in the solution. Rinse the substrate completely with PBS. ToF SIMS analysis was used to confirm the immobilization of the antibody on the PPOx substrate.

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PUM

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Abstract

A device for selective capture of target bladder cancer cells from urine or a urine derived fluid is provided. The device comprises a substrate having one or more cell capture surfaces, each cell capture surface comprising a functionalized film on the substrate and one or more target bladder cancer cell selective binding agents covalently bound to the functionalized film.

Description

[0001] Priority document [0002] This application claims the priority of Australian Provisional Patent Application No. 2017901350 entitled "BLADDER CANCER DETECTION DEVICEAND METHOD" filed on April 12, 2017, the contents of which are fully incorporated herein by reference. Technical field [0003] The present disclosure relates to devices and methods for selectively capturing and / or detecting bladder cancer cells, or their biomarkers, from urine. The device and method can be particularly applied to point of care diagnosis devices and methods. Background technique [0004] The bladder is the most common cancer site in the urinary tract and the most common histological type is transitional cell carcinoma. Patients with one type of bladder cancer are at high risk of developing other urinary tract cancers and have a particularly high recurrence rate (70%) (Dawam 2012; Youssef and Lotan 27 2011). Therefore, it is necessary to monitor patients with bladder cancer regularly and indefini...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/53
CPCG01N33/54366G01N33/56966G01N33/57407
Inventor 克拉西米尔·阿塔纳索夫·瓦西里夫梅勒妮·麦格雷戈乔纳森·格利德尔乔丹·李
Owner UNIVERSITY OF SOUTH AUSTRALIA
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