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Porous silicon-based non-labelling real-time online detection method for cholera toxins

A technology for cholera toxin and porous silicon, which is used in the measurement of phase-influenced properties, etc.

Inactive Publication Date: 2014-02-05
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of these techniques involve complex optical systems, temperature control systems or metal-enhanced surface layers

Method used

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  • Porous silicon-based non-labelling real-time online detection method for cholera toxins
  • Porous silicon-based non-labelling real-time online detection method for cholera toxins
  • Porous silicon-based non-labelling real-time online detection method for cholera toxins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Hydrophobicity and stability verification of the film of embodiment 1

[0041] (1) First, silicon wafers were prepared by electrochemical etching to prepare porous silicon. The etching solution was hydrofosic acid: absolute ethanol (volume ratio 3:1), the current sensitivity was 486mA / cm2, and the etching time was 10s.

[0042] (2) Hydrophobic modification of silicon surface by 1H, 1H, 2H-perfluoro-1-decene at 140°C under nitrogen protection for 2 hours. Hydrophobic modified silicon wafers were obtained.

[0043] (3) The hydrophobically modified silicon wafer is fixed on the bottom of the polycarbonate flow cell, and the Y-shaped optical fiber is perpendicular to the surface of the flow cell. The reflection interference spectrum is obtained through the optical fiber, and a data is recorded every 6 seconds. The Liye is transformed into a reflection peak spectrum, with time as the axis of abscissa and intensity of the reflection spectrum as the axis of ordinate, and is ...

Embodiment 2

[0048] Steps (1)-(3) are the same as in Example 1,

[0049] (4) Liposome-monosialoganglioside nano-mixture was prepared with a liposome extruder; the molar ratio of monosialoganglioside and liposome microspheres was 1:40, and extruded through a pore size of 50nm of polycarbonate film 21 times.

[0050] (5) Immobilization of liposomes and monosialoganglioside. Immobilization of liposome-monosialoganglioside on porous silicon surfaces via hydrophobic interactions. Specifically, the flow cell is first washed with deionized water for 15-20 minutes, then washed with pH7.4 phosphate buffer for 5 minutes, and then, 0.1mg / mL liposome and monosialoganglioside mixture through the flow cell until the optical thickness of the film reaches a stable value, see Figure 4 .

[0051] (6) Non-labeled on-line detection of cholera toxin: After washing the sample (containing cholera toxin solution) for 5 minutes with pH 7.4 phosphate buffer solution, the phosphate solution of 1nM cholera toxin...

Embodiment 3

[0053] Embodiment 3: Specific detection

[0054] Specificity detection 1: basically the same as Example 2, the difference is that the liposome microspheres without monosialoganglioside are immobilized on the hydrophobically modified silicon wafer (porous silicon film), and then according to the (6) For detection, the sample contains 20nM cholera toxin, see Image 6 . Depend on Image 6 It can be seen that after the introduction of 20nM cholera toxin, the optical thickness of the film does not red shift. This is because there is no receptor for cholera toxin on the surface of the film, and the cholera toxin is not bound to the surface of the film to cause a change in the refractive index of the film.

[0055] Specificity detection 2: basically the same as Example 2, the difference is that the sample is a phosphate buffer solution containing 20nM bovine serum albumin (without cholera toxin) circulating in the flow cell through a microflow pump, and monitoring the thin film opt...

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Abstract

The invention relates to a porous silicon-based non-labelling real-time online detection method for cholera toxins. The method includes: using porous silicon as a sensing substrate, modifying the porous silicon surface with hydrophobic groups, immobilizing liposome and monosialoganglioside via hydrophobic interaction, when a sample contains cholera toxins, combining monosialoganglioside (GM1) specifically with toxin molecules, making use of the reflection interference spectrum of porous silicon, performing Fourier infrared transformation, then conducting real-time non-labelling detection on porous silicon effective film thickness change, and carrying out quantitative and qualitative detection on cholera toxins. The method can realize real-time online detection of cholera toxins, the lowest detection limit is 1nM, and the sensing substrate can be reused.

Description

technical field [0001] The invention designs a rapid, low-cost, non-marker real-time on-line detection technology method for cholera toxin in food. Specifically, by hydrophobically modifying the surface of porous silicon, liposomes and monosialogangliosides are immobilized on the surface of silicon wafers through hydrophobic interactions, and using the reflection interference spectrum of porous silicon, after Fourier transform infrared transformation , real-time label-free detection of thickness changes in porous silicon effective films to detect cholera toxin in food. Background technique [0002] Cholera toxin is an intracellular toxin produced by Vibrio cholerae. Generally, it consists of A subunit with catalytic function and five B subunits with recognition function. If people and animals eat food or feed contaminated by cholera toxin, it will cause serious consequences such as poisoning and even death. The rapid quantitative detection of cholera toxin is of great sig...

Claims

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Application Information

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IPC IPC(8): G01N21/45
Inventor 李建林李玮徐坤曹斌孙越徐杰蒋云坤郑铁松
Owner NANJING NORMAL UNIVERSITY
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