Universal type multi-signal output biosensor, as well as preparation method and application thereof

A biosensor, multi-signal technology, applied in Raman scattering, color/spectral property measurement, material excitation analysis, etc., can solve the problems of limited color change detection sensitivity, unsuitable for detection, and no similar technology found, etc. Low impact, strong specificity and high efficiency

Inactive Publication Date: 2015-07-29
LUDONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the limited detection sensitivity of human eyes to color changes, this method is only suitable for qualitative and semi-quantitative detection of bacterial liquids with relatively high concentrations.
The advantage of SPR peak shift is that it has high

Method used

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  • Universal type multi-signal output biosensor, as well as preparation method and application thereof
  • Universal type multi-signal output biosensor, as well as preparation method and application thereof
  • Universal type multi-signal output biosensor, as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The preparation process of a general-purpose multi-signal output biosensor is as follows:

[0032] (1) Preparation of gold-silver composite nanoparticles with silver-coated gold core-shell structure:

[0033] Add 82.2 μL of 0.01% (mass volume percent concentration, W / V) chloroauric acid solution into 50 mL of ultrapure water, stir vigorously and heat to 100 °C. After reacting for 30 minutes, slowly add 500 μL of 1% (W / V) sodium citrate solution dropwise, and continue heating for 15 minutes. Then, 235 μL of 0.1 mol / L silver nitrate solution and 800 μL of 1% (W / V) sodium citrate solution were slowly added dropwise. After the reaction system was continued to be heated for 1 hr, an orange-red, silver-coated gold, and core-shell structure composite nanoparticle solution was obtained. The transmission electron microscope and photodynamic scattering diagrams of the gold-silver composite nanoparticles prepared in this example are shown in image 3 .

[0034] (2) Synthesis o...

Embodiment 2

[0040] Embodiment 2: The biosensor of universal multi-signal output detects Gram-positive pathogenic bacteria Enterococcus Enterococcus Faecalis (ATCC 51299).

[0041] (1) Gram-positive pathogenic bacteria Enterococcus were selected Enterococcus Faecalis As a model, the detection performance of the general-purpose multi-signal output biosensor is investigated, and the detection results are as follows: Figure 5 shown. exist Figure 5 (a) and (b) are the qualitative and quantitative detection results of chromaticity and ultraviolet-visible light spectroscopy, respectively; (c) and (d) are the qualitative and quantitative analysis results of surface-enhanced Raman spectroscopy.

[0042] (2) The general-purpose multi-signal output biosensor can realize the detection of Gram-positive pathogenic bacteria Enterococcus Enterococcus Faecalis qualitative and quantitative detection. First, the cultured bacteria were washed three times with PBS buffer and diluted to different con...

Embodiment 3

[0043] Example 3: Universal multi-signal output biosensor detects Gram-positive pathogenic bacteria Enterococcus Enterococcus Faecium (ATCC 51559).

[0044] Enterococcus Enterococcus Faecium As a model, the detection performance of the general-purpose multi-signal output biosensor was investigated. A general-purpose multi-signal output biosensor enables detection of Gram-positive pathogenic bacteria Enterococcus Enterococcus Faecium qualitative and quantitative detection. First, the cultured bacteria were washed three times with PBS buffer and diluted to different concentrations of the bacterial solution; secondly, 10 μL of 0.1 mmol / L trans-cyclooctene-modified vancomycin derivative was added to 90 μL of the bacterial solution of different concentrations Incubate at 37°C for 30 min; then, add 500 μL of functionalized gold-silver composite nanoparticle solution, incubate at 37°C for 10 min, observe the color change of the solution, and test the UV-visible spectrum and Ram...

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Abstract

The invention discloses a universal type multi-signal output biosensor, as well as a preparation method and application thereof. The universal type multi-signal output biosensor comprises two parts: a transcyclooctene modified vancomycin derivative used as a recognition molecule and functionalized gold and silver compound nanoparticles used as a signal conversion element, wherein the amino activated locus of the vancomycin molecule is covalently coupled with transcyclooctene; the gold and silver compound nanoparticles adopt silver shell-gold core compound nanoparticles; the surfaces of the gold and silver compound nanoparticles are simultaneously modified with a disulfide bond-containing tetrazine compound and a Raman microprobe molecule. The universal type multi-signal output biosensor can be applied to detection of Gram-positive pathogens, has the advantages of being high in efficiency, low in external environment influence, wide-spectral in targets and the like, and can meet the requirement of actual production and living.

Description

Technical field: [0001] The invention relates to a general-purpose multi-signal output biosensor and its preparation method and its application in the detection of Gram-positive pathogenic bacteria, belonging to the detection of pathogenic bacteria and the preparation technology of nanometer materials. Background technique: [0002] So far, the continuous spread of various pathogenic bacteria, especially Gram-positive pathogenic bacteria (such as Staphylococcus aureus, Streptococcus pneumoniae, Enterococcus, etc.), has become one of the most important causes of human death worldwide. one. In recent years, with the abuse of antibiotics, multidrug-resistant bacterial infections have gradually become a difficult problem in the international medical community, and have also become an important reason for prolonged hospitalization, increased medical expenses, and patient death in my country. Studies have shown that early detection of the presence of pathogenic bacteria can eff...

Claims

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Application Information

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IPC IPC(8): G01N21/25G01N21/31G01N21/65
Inventor 江婷婷王远芳王莹范华莹刘佩龙
Owner LUDONG UNIVERSITY
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