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Method for preparing fluorescently-encoded microsphere test strip for simultaneous detection of multiple staphylococcal enterotoxins

A staphylococcus intestinal tract and fluorescent coding technology, applied in the field of preparation of fluorescent coding microsphere test strips, can solve the problems of only colorimetric detection and analysis, cumbersome operation, high cost, etc., and achieve qualitative and quantitative detection, particle size Uniform distribution and easy operation

Active Publication Date: 2019-12-27
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These traditional methods are time-consuming, cumbersome, and costly, and are not suitable for rapid screening of food samples. Therefore, it is necessary to study a method for rapid detection of enterotoxins in food
[0003] Immunochromatographic detection technology is a rapid detection method developed by combining immunological technology of antigen-antibody immunological reaction and chromatography technology. The main labeling materials include radioactive materials, enzymes, colloidal gold and fluorescein. The main immunochromatographic test strips on the market are mainly colloidal gold-labeled immunochromatographic test strips, which can only realize the detection and analysis of one substance or colorimetric detection and analysis; with the rapid development of new nanomaterials, fluorescent Due to its stable morphological structure and stable and efficient luminous efficiency, encoded microspheres show great application potential in the fields of labeling, tracing, and detection; therefore, it is necessary to study a method for simultaneously detecting multiple Staphylococcus aureus enterotoxins Immunochromatographic test strips, in order to greatly improve the detection efficiency and detection accuracy of food samples

Method used

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  • Method for preparing fluorescently-encoded microsphere test strip for simultaneous detection of multiple staphylococcal enterotoxins
  • Method for preparing fluorescently-encoded microsphere test strip for simultaneous detection of multiple staphylococcal enterotoxins
  • Method for preparing fluorescently-encoded microsphere test strip for simultaneous detection of multiple staphylococcal enterotoxins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The present invention prepares a carbon quantum dot fluorescence-coded microsphere immunochromatographic test strip for simultaneously detecting three kinds of Staphylococcus aureus enterotoxins, the three kinds of Staphylococcus aureus enterotoxins are enterotoxin A, enterotoxin B and enterotoxin C , including the following steps:

[0054] Step 1, preparing carbon quantum dot fluorescence encoded microspheres;

[0055] First, mix 1g of sucrose and 10mL of phosphoric acid and heat them in a microwave oven (750W) for 1min and 30s, respectively, to obtain two kinds of fluorescent carbon quanta, which are denoted as CQD1 ​​and CQD2. The emission peaks are 440nm and 650nm, showing blue and red respectively. Fluorescence; according to 1:0, 3:1 and 0:1, CQD1 ​​and CQD2 were premixed in three different proportions, and 1mg poly(maleic anhydride-alt-1-octadecene), 1.2mg polymethyl Methyl acrylate and 0.5 mg of carbon quantum dots in a good proportion were dispersed in 10 mL of...

Embodiment 2

[0072] This implementation example prepares carbon quantum dot fluorescent coding microsphere immunochromatographic test strips and detects Staphylococcus aureus enterotoxin B (SEB), specifically including the following steps:

[0073] Step 1, preparing carbon quantum dot fluorescence encoded microspheres;

[0074] First, mix 1g of sucrose and 10mL of phosphoric acid and heat them in a microwave oven (750W) for 1min and 30s, respectively, to obtain two kinds of fluorescent carbon quanta, which are denoted as CQD1 ​​and CQD2. The emission peaks are 440nm and 650nm, showing blue and red respectively. Fluorescence: Premix CQD1 ​​and CQD2 according to a ratio of 1:1, and mix 1mg poly(maleic anhydride-alt-1-octadecene), 1.2mg polymethyl methacrylate and 0.5mg Carbon quantum dots were dispersed in 10mL chloroform; then 0.3mg sodium dodecylsulfonate was added to the above chloroform dispersion for ultrasonic dispersion for 10min; then 0.2mg sodium hydroxide was added for ultrasonicat...

Embodiment 3

[0088] Example 3 of the present invention prepares carbon quantum dot fluorescence-encoded microsphere immunochromatographic test strips for simultaneous detection of 6 kinds of Staphylococcus aureus enterotoxins, the 6 kinds of Staphylococcus aureus enterotoxins are enterotoxins A, B, C, D, E, and F, the specific steps include:

[0089] Step 1, preparing carbon quantum dot fluorescence encoded microspheres;

[0090] First, mix 1g of sucrose and 10mL of phosphoric acid and heat them in a microwave oven (750W) for 1min and 30s, respectively, to obtain two kinds of fluorescent carbon quanta, which are denoted as CQD1 ​​and CQD2. The emission peaks are 440nm and 650nm, showing blue and red respectively. Fluorescence; according to 1:0, 1:2, 1:3, 0:1, 2:1 and 3:1, premix CQD1 ​​and CQD2 in 6 different ratios, and mix 1mg poly(maleic anhydride-alt-1 -octadecene), 1.2mg polymethyl methacrylate and 0.5mg carbon quantum dots with a good ratio are dispersed in 10mL chloroform; then 0.3...

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Abstract

The invention belongs to the field of rapid food safety detection, and relates to a method for preparing a fluorescently-coded microsphere test strip for simultaneous detection of multiple staphylococcal enterotoxins, and the method comprises the following steps of firstly preparing two fluorescent carbon quantum dots with good fluorescent characteristics, good water solubility and good biocompatibility, and preparing a series of fluorescently-coded microspheres with different fluorescence characteristics by adjusting the doping ratio of the two carbon quantum dots; fluorescently coding microspheres by the carbon quantum dots, and correspondingly labeling the antibody of enterotoxins to obtain a plurality of fluorescent antibody probes; finally dissolving the plurality of fluorescent antibody probes in a phosphate buffer solution containing sucrose and Tween 20 at equal quality to obtain a mixed solution containing the plurality of fluorescent antibody probes; and evenly spraying the mixed solution onto the glass fiber membrane as a bonding pad to prepare the fluorescent microsphere immunochromatographic test strip, so as to realize the simultaneous qualitative and quantitative detection of multiple enterotoxins. The method provided by the invention can improve the detection efficiency and accuracy and achieve rapid detection of mass food samples.

Description

technical field [0001] The invention belongs to the field of rapid detection of food safety, and in particular relates to a preparation method of a fluorescent coded microsphere test strip for simultaneously detecting multiple Staphylococcus aureus enterotoxins. Background technique [0002] Staphylococcus aureus is a common food-borne pathogen that widely exists in nature. Staphylococcal enterotoxin (SE) secreted by Staphylococcus aureus is the main cause of food poisoning caused by Staphylococcus aureus. Studies have shown that most Staphylococcus aureus can produce various enterotoxins such as A, B, C, D, E and F. In the process of food processing, the enterotoxin produced by the exocrine secretion of bacteria is extremely resistant to heat, and cannot be completely destroyed when heated to 100°C for 30 minutes. After ingestion, it can still resist the decomposition of protease in the stomach and cause Human vomiting, diarrhea and other symptoms. The current methods for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/569G01N33/558G01N33/543
CPCG01N33/56938G01N33/582G01N33/585G01N33/558G01N33/54313
Inventor 邹小波胡雪桃石吉勇李艳肖甘子玉徐艺伟李亚惠
Owner JIANGSU UNIV
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