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Two-dimensional molecular engram array type quality sensing detection method for staphylococcus enterotoxin

A staphylococcal intestinal and molecular imprinting technology, which is used in measurement devices, analytical materials, material analysis by electromagnetic means, etc., can solve the problem of poor substance transport and permanent template retention and recognition performance, and is not suitable for identifying macromolecular proteins and macromolecules. The template is difficult to achieve and other problems, to achieve the effect of rapid detection, simple preparation method and low cost

Inactive Publication Date: 2013-09-25
INST OF HYGIENE & ENVIRONMENTAL MEDICINE PLA ACAD OF MILITARY MEDICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, traditional imprinted polymers are three-dimensional blocks with high density, and it is difficult for macromolecular templates to reach and leave the binding site. Poor material transport and permanent template retention have a negative impact on the recognition performance. Imprinting conditions and organic solvents commonly used in experiments will denature proteins, making three-dimensional imprinting techniques unsuitable for identifying macromolecular proteins

Method used

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  • Two-dimensional molecular engram array type quality sensing detection method for staphylococcus enterotoxin
  • Two-dimensional molecular engram array type quality sensing detection method for staphylococcus enterotoxin
  • Two-dimensional molecular engram array type quality sensing detection method for staphylococcus enterotoxin

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Experimental program
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Effect test

Embodiment 1

[0046] A two-dimensional molecular imprinted array mass sensing detection method for multiple Staphylococcus aureus enterotoxins in food, comprising the following steps:

[0047] (1) Surface active treatment of quartz crystal oscillator electrodes; immerse three gold-plated quartz crystal oscillator electrodes in a 0.1M ethanol solution of 11-mercaptoalcohol acid, soak for 12 hours, take them out, rinse with absolute ethanol, blow dry with nitrogen, and mark them respectively For A, B and C electrodes;

[0048] (2) Preparation of initial sol-gel:

[0049] Mix 2.5 mL of tetraethylorthosilicate, 150 μl of phenyltrimethoxysilane, 150 μl of methyltrimethoxysilane, 500 μl of 0.1 M aqueous HCl and 2 mL of H 2 O mixed, ultrasonic 25 minutes is prepared into initial sol-gel;

[0050] (3) Preparation of imprinted sol-gel containing templates of SEA or SEB:

[0051] Take 3 mL of the initial sol-gel and 150 μl of SEA phosphate buffer with a concentration of 0.1 mg / mL, mix well, and so...

Embodiment 2

[0067] A two-dimensional molecular imprinted array mass sensing detection method for multiple Staphylococcus aureus enterotoxins in food, comprising the following steps:

[0068] (1) Surface active treatment of quartz crystal oscillator electrodes; immerse three gold-plated quartz crystal oscillator electrodes in a 0.05M ethanol solution of 11-mercaptoalcohol acid, soak for 16 hours, take them out, rinse them with absolute ethanol, dry them with nitrogen, and mark them respectively For A, B and C electrodes;

[0069] (2) Preparation of initial sol-gel:

[0070] Mix 2 mL of tetraethylorthosilicate, 200 μl of phenyltrimethoxysilane, 100 μl of methyltrimethoxysilane, 600 μl of 0.05 M aqueous HCl and 3 mL of H 2 O mixed, ultrasonic 20 minutes is prepared into initial sol-gel;

[0071] (3) Preparation of imprinted sol-gel containing templates of SEA or SEB:

[0072] Take 2 mL of initial sol-gel and 100 μl of SEA phosphate buffer with a concentration of 0.2 mg / mL, mix well, and s...

Embodiment 3

[0086] A two-dimensional molecular imprinted array mass sensing detection method for multiple Staphylococcus aureus enterotoxins in food, comprising the following steps:

[0087] (1) Surface active treatment of quartz crystal oscillator electrodes; immerse three gold-plated quartz crystal oscillator electrodes in a 0.2M ethanol solution of 11-mercaptoalcohol acid, soak for 8 hours, take it out, rinse with absolute ethanol, blow dry with nitrogen, and mark them separately For A, B and C electrodes;

[0088] (2) Preparation of initial sol-gel:

[0089] Mix 3 mL of tetraethylorthosilicate, 100 μl of phenyltrimethoxysilane, 200 μl of methyltrimethoxysilane, 400 μl of 0.2 M aqueous HCl and 2 ml of H 2 O mixed, ultrasonic 30 minutes is prepared into initial sol-gel;

[0090] (3) Preparation of imprinted sol-gel containing SEA or SEB template:

[0091] Take 4 mL of initial sol-gel and 200 μl of SEA phosphate buffer with a concentration of 0.3 mg / mL, mix well, and sonicate for 40 m...

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Abstract

The invention discloses a two-dimensional molecular engram array type quality sensing detection method for staphylococcus enterotoxin, which comprises the following steps: (1) performing surface active treatment on a quartz oscillating electrode; (2) preparing an initial sol-gel; (3) preparing an engram sol-gel of a template containing SEA (staphylococcus enterotoxin A) and SEB (staphylococcus enterotoxin B); (4) performing coating treatment on the surface of the quartz oscillating electrode; (5) removing protein molecules SEA and SEB of the template; and (6) constructing a molecular engram array quality sensor for the SEA and SEB detection. The conventional SEA and SEB macromolecular proteins in the staphylococcus enterotoxin which is the important virulence factor of sitotoxismus are quickly detected. The method is simple; no mark is required; the detection speed is high; the external interference is small; the cost is low; the result is stable and reliable; the time of the whole detecting process is short; and the method has a wide application prospect in the fields of food safety, hygiene detection, environment monitoring, clinical examination, biological terror monitoring, and the like.

Description

technical field [0001] The invention relates to a detection method of Staphylococcus aureus enterotoxin in food, in particular to a detection method of Staphylococcus enterotoxin A (SEA) and B (SEB). Background technique [0002] Staphylococcus aureus is a common food poisoning pathogen, which can cause various diseases in humans and animals. Its pathogenicity is mainly closely related to the enterotoxin it secretes. Staphylococcal enterotoxins (SEs) are a group of heat-resistant low-molecular-weight soluble proteins, among which SEB is an important pathogenic factor of food poisoning and is listed as one of the main toxins of biological warfare agents. SEB protein is stable to heat, and the lowest concentration of causing food poisoning is 200ng / 100g of food. People who accidentally eat food contaminated by it will cause nausea, vomiting and diarrhea after 4-6 hours. Therefore, research on rapid and sensitive detection of SEB and other SEs such as SEA, SEC 1 , SEC 2 The ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/333
Inventor 刘楠高志贤马新华欧国荣李晓丽陈翔
Owner INST OF HYGIENE & ENVIRONMENTAL MEDICINE PLA ACAD OF MILITARY MEDICAL