Surface plasmon resonance sensor chip for detecting gram-negative bacteria and preparation method and application thereof

A technology of surface plasma and Gram-negative bacteria, which is applied in measuring devices, material analysis through optical means, instruments, etc., to achieve the effects of high sensitivity, simple operation steps and clear structure

Active Publication Date: 2018-06-15
TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no SPR sensor suitable for detecting Gram-negative bacteria, and SPR technology has not been used to detect Gram-negative bacteria in aqueous solutions in real time, quickly, easily, quantitatively, and ultra-sensitively, especially in water for injection. Gram-negative bacteria content method

Method used

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  • Surface plasmon resonance sensor chip for detecting gram-negative bacteria and preparation method and application thereof
  • Surface plasmon resonance sensor chip for detecting gram-negative bacteria and preparation method and application thereof
  • Surface plasmon resonance sensor chip for detecting gram-negative bacteria and preparation method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0053] A preparation method for Gram-negative bacteria detection surface plasmon resonance sensor chip, comprising the following steps:

[0054] (1) Gold is plated on the surface of the glass substrate, and the thickness of the gold film is 40nm;

[0055] (2) Soak the glass substrate obtained in step (1) in the compound S1 solution of 0.01mmol / L, let it stand for 1 hour, rinse it with water repeatedly, and set aside; wherein, the structure of compound S1 is:

[0056]

[0057] In the formula, Z and R are ester groups and amino groups respectively; the solvent of the compound S1 solution is physiological saline.

[0058] (3) Streptomycin sulfate, kanamycin sulfate and gentamycin sulfate probe molecules, NHS, EDC, HEPES buffer are mixed according to the molar ratio of 1:10:10:1 to obtain a mixed solution; (2) The obtained glass substrate is soaked in the mixed solution and left to stand for 2 hours;

[0059] (4) Take out the glass substrate, wash the glass substrate repeated...

Embodiment 2

[0061] A preparation method for Gram-negative bacteria detection surface plasmon resonance sensor chip, comprising the following steps:

[0062] (1) Gold is plated on the surface of the glass substrate, and the thickness of the gold film is 60nm;

[0063] (2) Soak the glass substrate obtained in step (1) in the compound S1-1 solution of 1000mmol / L, let it stand for 24 hours, rinse it repeatedly with distilled water, and set aside; wherein, the structure of compound S1-1 is:

[0064]

[0065] In the formula, the solvent of the compound S1-1 solution is acetonitrile

[0066] (3) Mix gentamicin sulfate probe molecules, NHS, EDC, and ethanol in a molar ratio of 1:100:100:1000 to obtain a mixed solution; soak the glass substrate obtained in step (2) in the mixed solution , stand still for 24 hours;

[0067] (4) Take out the glass substrate, rinse the glass substrate repeatedly with ethanol and distilled water, and obtain a surface plasmon resonance sensor chip for Gram-negativ...

Embodiment 3

[0069] A preparation method for Gram-negative bacteria detection surface plasmon resonance sensor chip, comprising the following steps:

[0070] (1) Gold is plated on the surface of the glass substrate, and the thickness of the gold film is 50nm;

[0071] (2) Soak the glass substrate obtained in step (1) in the compound S1-2 solution of 500mmol / L, let it stand for 12 hours, rinse it repeatedly with distilled water, and set aside;

[0072] Wherein, the structure of compound S1-2 is:

[0073]

[0074] The solvent of the compound S1-2 solution is chloroform;

[0075] (3) Mix the Genda-Pionomycin probe molecule, NHS, EDC, N,N-dimethylacetamide according to the molar ratio of 1:10:10:100 to obtain a mixed solution; the step (2) The obtained glass substrate was soaked in the mixed solution and left to stand for 12 hours;

[0076] (4) Take out the glass substrate, rinse the glass substrate repeatedly with ethanol and distilled water, and obtain a surface plasmon resonance senso...

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Abstract

The invention discloses a surface plasmon resonance sensor chip for detecting gram-negative bacteria and a preparation method and application thereof. The chip comprises a glass substrate layer, a gold film layer and a probe molecular layer. The gold film layer is disposed on the glass substrate layer. The probe molecular layer is disposed on the gold film layer. A probe molecule of the probe molecule layer contains at least one of drug molecules of streptomycin sulfate, kanamycin sulfate, gentamicin sulfate, gentamycin-micronomicin, vistamycin sulfate, neomycin, polymyxin B, rifampicin, chloramphenicol and fosfomycin. The invention also discloses a preparation method and application of the chip. The surface plasmon resonance sensor chip creatively realizes detection of gram-negative bacteria based on surface plasmon resonance, has high sensitivity and good selectivity, has a gram-negative bacterium concentration detection linear range of 10-10<6> CFU/mL and can be used for detecting gram-negative bacteria in solutions such as detection water, drinking water or human body fluids.

Description

technical field [0001] The invention relates to the field of surface plasmon sensor chip preparation. More specifically, it relates to a surface plasmon resonance sensor chip that can be used for the detection of Gram-negative bacteria and its preparation method and application. Background technique [0002] Bacteria are both useful and harmful to the environment, humans and animals. Some bacteria became pathogens, causing tetanus, typhoid, pneumonia, syphilis, cholera, and tuberculosis. In plants, the bacteria cause leaf spot, fire blight and wilting. Modes of infection include contact, airborne transmission, food, water, and carrier microorganisms. Bacteria also have a great impact on human activities. On the one hand, bacteria are the pathogens of many diseases, including tuberculosis, gonorrhea, anthrax, syphilis, plague, trachoma and other diseases are caused by bacteria. The significance of the Gram staining method is to identify bacteria and divide many bacteria ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/552
CPCG01N21/553
Inventor 张洪艳李亚思刘卫敏葛介超吴加胜汪鹏飞
Owner TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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