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Biological chip for rapidly screening bladder cancer markers as well as preparation method and application thereof

A biochip and bladder cancer technology, applied in the field of analysis and detection, can solve the problems of unsuitability for early screening and diagnosis of bladder cancer, strong invasiveness of bladder cancer examination, and low diagnostic efficiency, and achieve easy integration and miniaturization, good electrical Features and stability, rapid detection of effects

Inactive Publication Date: 2021-10-01
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, cystoscopy is the gold standard method for clinical diagnosis of bladder cancer, but this technique is limited by the volume, location and environment of the cancer, resulting in low resolution, and bladder cancer examination is highly invasive, and patients may experience urethral pain and hematuria, etc. problem, not suitable for early screening and diagnosis of bladder cancer
Although urine exfoliation cytology examination and imaging examination are not invasive, their diagnostic efficiency is low, and they are not suitable for early screening and diagnosis of bladder cancer
Therefore, it is still a severe challenge in the rapid diagnosis and early screening of bladder cancer.

Method used

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  • Biological chip for rapidly screening bladder cancer markers as well as preparation method and application thereof
  • Biological chip for rapidly screening bladder cancer markers as well as preparation method and application thereof
  • Biological chip for rapidly screening bladder cancer markers as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: the preparation of nano biochip

[0040] S1. Preparation of metal source and drain electrodes

[0041] S1.1) Select a p-type silicon wafer with a silicon oxide layer of 300nm as the substrate, spin-coat a layer of positive photoresist AZ521 on its surface, heat it on a heating plate at 110°C for 5min, and then use a UV photolithography machine on the mask Under the protection of the stencil pattern, the electrode channel is engraved, developed in the ZX-238 developer for 50s, and the silicon wafer is rinsed with deionized water to remove the residual developer.

[0042] S1.2) A Cr / Au electrode is grown by evaporation using a thermal evaporation apparatus, wherein the Cr layer is on the bottom with a thickness of 10 nm, and the Au layer is on the top with a thickness of 50 nm. Metal Cr can increase the bonding force between the oxide layer of the silicon wafer and the metal Au. Soak the silicon wafer deposited with electrodes in an acetone solution to peel...

Embodiment 2

[0057] Embodiment 2: the determination of cDNA / IGZO TFT device standard curve

[0058] The cDNA / IGZO TFT device prepared in Example 1 is tested to RNA solutions with different concentration gradients, and the source-drain electrode voltage V d unchanged (V d =0.1V), by changing the gate voltage V g To obtain the electrical signal change curve of the device to RNA solutions with different concentration gradients. By continuously changing the concentration gradient of the RNA solution, the electrical signal change curve of the device to a series of miRNA concentrations was measured. From Figure 4 As shown, with the increasing concentration of RNA, more and more RNA molecules will be bound on the surface of the cDNA / IGZO TFT device. At this time, the negative gate effect exerted by the negatively charged RNA chain will become larger and larger, resulting in a gap in the device. The carrier concentration in the channel gradually decreases, and the current decreases gradually....

Embodiment 3

[0059] Example 3: Extraction of nucleic acid marker miRNA in patient's urine

[0060] 1. Take 0.5mL of the patient's urine sample, add 1.5mL of lysis buffer (Lysis buffer), and blow the urine sample with a pipette gun to speed up the lysis of cells in the urine sample. (The volume ratio of Lysis buffer to urine sample is always 3:1)

[0061] 2. Shake the sample vigorously and incubate at 15-30°C for 5 minutes to completely decompose the ribosomes.

[0062] 3. Add 0.2mL chloroform to the mixed solution, shake vigorously for 15s and place at room temperature for 2min.

[0063] 4. Centrifuge the mixed solution at 4°C, 12,000rpm for 10min, the sample will be divided into three layers: the lower layer is the organic phase, the middle layer and the upper layer are colorless aqueous phases, in which RNA exists in the aqueous phase.

[0064] 5. Take the supernatant and transfer it to a new centrifuge tube, add 1.5 times the volume of absolute ethanol at the same time, and vortex eve...

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Abstract

The invention discloses a biological chip for rapidly screening bladder cancer markers as well as a preparation method and application thereof, and belongs to the field of analysis and detection. According to the nanometer biochip, the corresponding linear relation between electric signals of microRNA (miRNA) and indium gallium zinc oxide (IGZO) sensing devices with different concentration gradients is established, the electric signal change of a urine sample of a bladder cancer patient is measured, and miRNA detection in an unknown urine sample is achieved. According to the nano biochip, the experimental process is greatly simplified, the requirement on experimental conditions is not high, an unknown urine sample can be rapidly detected, and the defects in the current detection field are well overcome. The nano biochip has the advantages of high sensitivity, high specificity, high detection speed, high repeatability and the like, and is very suitable for early screening of bladder cancer.

Description

technical field [0001] The invention relates to the field of analysis and detection, in particular to a biochip for rapid screening of bladder cancer markers, a preparation method and application thereof. Background technique [0002] Bladder cancer is the most common malignant tumor in the urinary system, with a high incidence rate and easy recurrence after surgery. At present, the clinical diagnosis of bladder cancer is mainly based on cystoscopy, but cystoscopy is highly invasive, has low resolution, and can easily cause urethral pain and hematuria during the inspection process, so it is difficult to be used for early screening of bladder cancer. As a urine storage organ, early cancer of the bladder will have a direct impact on the composition of the patient's urine. Currently, researchers have found multiple types of bladder cancer biomarkers in urine, including DNA, RNA, protein, and tumor cells. However, the concentration of these markers in urine is extremely low, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26
CPCG01N27/26
Inventor 袁荃杨雁冰王敬锋
Owner WUHAN UNIV
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