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Preparation method of PAMAM silicon-based cascade double-ring resonant cavity chip for detecting PCA3

A double-ring resonance and cascading technology, which is applied in the direction of color/spectral characteristic measurement, etc., can solve the problems of not observing the signal changes of cascaded double-ring resonant cavity sensors, the inability to realize PCA3 selective detection, and the small number of PCA3 gene probes, etc. , to increase the quantity, reduce the detection cost, enhance the signal strength and sensitivity

Inactive Publication Date: 2021-05-11
济南国科医工科技发展有限公司
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Problems solved by technology

[0006] The silicon-based cascaded double-ring resonant cavity sensor has high refractive index sensitivity, which can realize the label-free detection of substances, but because the cascaded double-ring resonant cavity sensor will show the same spectral signal for substances with the same refractive index, it cannot be realized. Selective detection of PCA3
If the method of directly connecting the PCA3 gene probe with the hydroxyl functional group on the surface of the resonator is used, the number of immobilized PCA3 gene probes is usually too small, so that the cascaded double-ring resonator cannot be observed when detecting PCA3 A change in the sensor signal, or a very small change in signal observed

Method used

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  • Preparation method of PAMAM silicon-based cascade double-ring resonant cavity chip for detecting PCA3

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Embodiment 1

[0066] Clean the silicon-based cascaded double-ring resonator with acetone, ethanol, and deionized water in sequence for 15 minutes, and then perform plasma treatment for 5 minutes at a power of 10.2w and a vacuum of 50pa (about 375mTor). The sensing ring of the silicon-based cascaded double-ring resonator was treated with 95% ethanol containing 2% (volume percentage) 3-(2,3-epoxypropoxy)propyltrimethoxysilane dropwise for 1 h, and then the Put the sensing ring into 15ml of DMSO solution containing 0.5g of G3PAMAM, and react at 100°C for 4h. Next, the sensing ring was put into 10 ml of DMSO solution containing 2.5 g of succinic anhydride and 0.15 g of DMAP, which was blown with nitrogen, and reacted at room temperature for 24 h. Then transfer the sensing loop to the newly prepared 0.04MEDC / 0.01M NHS solution and let it stand for 1h, then transfer to the NH solution containing 1μM PCA3 probe. 2 -(CH 2 ) 6 -AAAAATTGTTCAAAGACCCTTCGTGTT was reacted overnight in a PBS solution a...

Embodiment 2

[0068] Clean the silicon-based cascaded double-ring resonator with acetone, ethanol, and deionized water in sequence for 15 minutes, and then perform plasma treatment for 5 minutes at a power of 10.2w and a vacuum of 50pa (about 375mTor). The surface of the sensing ring of the silicon-based cascaded double-ring resonator was treated with anhydrous toluene solution containing 1% (volume percentage) of 3-(2,3-epoxypropoxy)propyltriethoxysilane for 30 minutes, and then After drying at 110° C. for 1 h, the sensing loop was put into 5 ml of methanol solution containing 10% (mass percent) G4 PAMAM, and reacted at 100° C. for 10 h. Then put the sensing loop into 10ml of nitrogen gas containing 30mM NaIO 4 The solution was reacted for 2h. Then the sensing loop was transferred to a freshly prepared 1% glutaraldehyde solution for 1 h, and then transferred to 1 μM PCA3 probe NH 2 -(CH 2 ) 6 -AAAAACTGCTGACTTTACCATCTGAGGCCAC was reacted overnight in a PBS solution at 4° C. to prepare a...

Embodiment 3

[0070]Clean the silicon-based cascaded double-ring resonator with acetone, ethanol, and deionized water in sequence for 15 minutes, and then perform plasma treatment for 5 minutes at a power of 10.2w and a vacuum of 50pa (about 375mTor). Add 10% (volume percent) 3-aminopropyltriethoxysilane aqueous solution (pH3-4) dropwise to the sensing ring of the silicon-based cascaded double-ring resonator and treat it at 75°C for 1 hour, then put the sensing ring into 5% glutaraldehyde solution at room temperature for 0.5 hours. After washing with 0.01MPBS (0.137MNaCl, pH7.4) buffer solution, the sensing loop was put into 5ml aqueous solution containing 150mg G5 PAMAM, and reacted at room temperature for 2h. Add 0.3 g of sodium borohydride and stir at room temperature for 36 hours. Next, the sensing ring was put into 4 ml of THF solution containing 51.6 mg of succinic anhydride and 15.7 mg of DMAP which was blown with nitrogen, and reacted at 40° C. for 4 h. Then transfer the sensing l...

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Abstract

The invention discloses a preparation method of a PAMAM silicon-based cascade double-ring resonant cavity chip for detecting PCA3, and belongs to the technical field of cascade double-ring resonant cavity sensing. The surface of a sensing ring in a silicon-based cascade double-ring resonant cavity is sequentially modified with a silane coupling agent, a PAMAM film and a PCA3 probe, and the silicon-based cascade double-ring resonant cavity chip for detecting PCA3 is prepared. Through the dendritic polyamino group in the PAMAM, the fixed number of PCA3 probes is increased, the specific binding number of the chip to PCA3 is increased, and the response signal of the silicon-based cascade double-ring resonant cavity sensor to PCA3 is enhanced. By utilizing the PAMAM silicon-based cascade double-ring resonant cavity chip with the modified PCA3 probe, the high-sensitivity and high-selectivity detection on the PCA3 can be realized. The chip disclosed by the invention does not need to be marked by fluorescent or luminous groups, so that the detection cost is saved, and the chip is expected to be popularized and applied in diagnosis and treatment of prostatic cancer.

Description

technical field [0001] The invention relates to the technical field of cascaded double-ring resonant cavity sensing, in particular to a method for preparing a PAMAM silicon-based cascaded double-ring resonant cavity chip for detecting PCA3. Background technique [0002] In recent years, the incidence of prostate cancer has gradually increased, and it has become the main malignant tumor that endangers men's health. Detecting the content of PSA in serum is the main screening method and auxiliary diagnostic index for prostate cancer. However, due to the limited sensitivity and specificity of PSA, this screening has low effectiveness and poor repeatability. The shortcomings of overdiagnosis and underdiagnosis of high-grade prostate cancer, and this kind of puncture biopsy is invasive, the patient suffers a lot, and it is easy to over-puncture, which can cause complications such as hematuria and rectal bleeding. [0003] The ideal prostate screening technique should be non-invas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/25
CPCG01N21/25
Inventor 李秋顺董文飞李力常智敏从瑛哥姜琛昱葛明锋宁珊珊
Owner 济南国科医工科技发展有限公司
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