Staphylococcal enterotoxin detection method

A staphylococcal enterotoxin detection method technology, which is applied in the field of rapid and high-sensitivity staphylococcal enterotoxin detection, can solve the problems of long detection time and achieve high sensitivity, short detection time and wide application range

Inactive Publication Date: 2009-08-26
INST OF OPTICS & ELECTRONICS - CHINESE ACAD OF SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0003] The problem to be solved by the present invention is: to overcome the shortcomings of the existing staphylococcal enterotoxin detection that requires special equipment and long detection time, and utilize LSRP (localized surface plasmon technology) and specific reactions between biochemical molecules to achieve Rapid, highly sensitive, label-free detection of staphylococcal enterotoxins

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Embodiment 1

[0034] In this embodiment, a detection chip with a metal silver structure with a characteristic size of 1000 nm on a quartz material substrate is used, and the detection method for staphylococcal enterotoxin of the present invention is used to detect SEB with a concentration of 10 μg / ml.

[0035] First, use quartz material as the chip substrate, clean it and perform hydrophilic treatment; make a cylindrical structure with an area of ​​about 10mm×10mm, a characteristic size of 1000nm, and an array number of 10×10 on the surface of the substrate by photolithography; through gold plating, the Nanostructure metallization to obtain a chip with nanogold structures on the surface of the substrate. Then add 2mM dimercaptodiacetic acid aqueous solution dropwise on the surface of the sample, and react for 20min; add 0.4M ethylcarbodiimide hydrochloride EDC and 0.1M hydroxysulfosuccinimide NHS mixed solution dropwise On the surface of the detection chip, activate the carboxyl group for 2...

Embodiment 2

[0037] In this embodiment, a detection chip with a gold-silver composite structure with a characteristic size of 500 nm is used on a germanium material substrate, and the detection method for staphylococcal enterotoxin of the present invention is used to detect SEB with a concentration of 0.1 μg / ml.

[0038] First, germanium material is used as the chip substrate, cleaned and subjected to hydrophilic treatment; a five-pointed star structure with an area of ​​about 500μm×500μm, a characteristic size of 500nm, and an array number of 4×4 is fabricated on the surface of the substrate by nanoimprinting; Silver metallizes the nanostructure to obtain a chip with a nanosilver structure on the surface of the substrate; then, a layer of 10nm thick gold is evaporated on the structure to form a double-layer composite structure. Then add 2mM dimercaptodiacetic acid aqueous solution dropwise on the surface of the sample, and react for 20min; add 0.4M ethylcarbodiimide hydrochloride EDC and 0...

Embodiment 3

[0040] In this embodiment, a detection chip with a gold structure with a characteristic size of 30 nm is used on a silicon material substrate, and the detection method for staphylococcus enterotoxin of the present invention is used to detect SEB with a concentration of 1 ng / ml.

[0041]First, silicon material is used as the chip substrate, cleaned and treated for hydrophilicity; on the surface of the substrate, diamond-shaped nanometers with an area of ​​about 20 μm × 20 μm, a characteristic size of 30 nm, and a quadrilateral arrangement of 1 × 1 are fabricated by self-assembly of nanospheres. structure; the nanostructure is metallized by silver plating to obtain a chip with a nanometal structure on the surface of the substrate. Then add 2mM dimercaptodiacetic acid aqueous solution dropwise on the sample surface and react for 20min; add 0.4M ethylcarbodiacin hydrochloride EDC and 0.1M hydroxysulfosuccinimide NHS mixed solution dropwise On the surface of the detection chip, act...

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Abstract

A staphylococcal enterotoxin detection method is characterized in that the detection method comprises the steps as follows: (1) preparing an LSPR detection chip of staphylococcal enterotoxin; (2) activating the surface of the metal structure of the chip to form a specific biological molecular membrane; (3) testing the extinction spectrum of the chip to obtain a reference value before combination; and (4) dripping a to-be-detected sample, putting the sample in an LSPR sensor for detection after the full reaction of the sample, observing spectral shift condition by using the specific reaction among antigen-antibody molecules, and judging whether the to-be-detected sample contains staphylococcal enterotoxin, so as to realize fast detection with high sensitivity and without a label. The method does not need complex equipment and does not use radioactive isotope, enzyme or fluorescence and the like as a label, and has the remarkable characteristics of quickness, high sensitivity, wide application range, security and high stability. The method provides a simple and practical new method for the quick detection of staphylococcal enterotoxin.

Description

technical field [0001] The invention belongs to the field of biochemical detection, relates to a novel staphylococcal enterotoxin detection method, in particular to a rapid and high-sensitivity staphylococcal enterotoxin detection method utilizing LSPR technology. Background technique [0002] The rapid detection and determination of the types of biological pathogens is the main purpose of the development of detection methods in various countries. In the face of various types of biological and chemical warfare agents, detection methods based on various principles have come out one after another. Detection method. Taking Staphylococcal enterotoxin type B (SEB) as an example, it is one of the Staphylococcus aureus enterotoxin series (SEA-SEE), which has strong heat resistance and drug resistance. The detection method mainly includes enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), etc. ELISA is the most commonly used method for detecting SEB. It ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N21/17
Inventor 邓启凌杜春雷罗先刚朱少丽杨欢史立芳
Owner INST OF OPTICS & ELECTRONICS - CHINESE ACAD OF SCI
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