Method of quantitatively detecting Listeria monocytogenes
A technology for quantitative detection of Listeria monocytogenes, which is applied in the field of analysis and detection, can solve the problems of hindering the application of immunochromatography technology and the low sensitivity of mobile rate immunochromatography technology, and achieves the prevention of false positive results, short detection time, and solution The effect of low sensitivity
Inactive Publication Date: 2019-11-19
QILU UNIV OF TECH
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[0005] In order to solve this bottleneck that hinders the application of immunochromatography technology, in the present invention, we have overcome the problem by replacing the classically used colloidal gold with magnetic gold nanoparticles, and using an external magnetic field to reduce its moving rate on the nitrocellulose membrane. The defect of low sensitivity of immunochromatography
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[0017] A method for quantitatively detecting Listeria monocytogenes, comprising the following steps:
[0018] 1) Preparation and modification of metallomimetic enzymes:
[0019] Preparation of seed solution: First, 5 mL of 0.2M cetyltrimethylammonium bromide solution and 5 mL of 0.5 mM tetrachloroauric acid solution were thoroughly mixed. Then, add 0.6mL 0.01M sodium borohydride solution, stir rapidly for 5 minutes and keep at 25°C for 1 hour;
[0020] Preparation of AuNR: First, 5 mL of 0.2M cetyltrimethylammonium bromide solution, 5 mL of 0.5 mM tetrachloroauric acid solution and 4.75 mL of deionized water were thoroughly mixed. Then, add 0.24mL 0.05M ascorbic acid solution and 2.5mL 4mM silver nitrate solution to the mixed solution, and stir vigorously at 30°C until the color of the solution changes from yellow to colorless (about 10 minutes). Finally, 20 μL of the seed solution prepared in ① was added, stirred for 30 seconds, and aged at 30°C for 12 hours;
[0021] ...
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Abstract
The invention relates to a method of quantitatively detecting Listeria monocytogenes, which includes the following steps: (1) a test strip of AuNR@Pt modified with 4-MBA and gold magnetic nanoparticles Fe3O4@Au is prepared; (2) a magnet is placed under the detection area of the test strip; and (3) the content of Listeria monocytogenes in a sample is determined by a color method and an SERS method. By replacing the classically used colloidal gold with magnetic gold nanoparticles and using a mode of reducing the moving rate on a nitro fiber membrane with an external magnetic field, the shortcoming of the low sensitivity of an immunochromatographic technology is overcome. The method combines the high selectivity, the enrichment and the flow rate controllability of the magnetic nanoparticleswith the high-throughput, fast, and simple advantages of the immunochromatographic technology, which greatly improves the detection sensitivity.
Description
technical field [0001] The invention belongs to the field of analysis and detection, and in particular relates to a method for quantitative detection of Listeria monocytogenes. Background technique [0002] Listeria monocytogenes, a Gram-positive short bacillus, is one of the main pathogens threatening human health. Because it widely exists in water bodies, public places, animal husbandry products, vegetables and fruits and other environments, the diseases caused by it often have a large number of people and are widely distributed, which poses a huge threat to food safety and people's health. Therefore, the detection of Listeria monocytogenes in food is of great significance. [0003] At present, the commonly used methods for detecting food-borne pathogens mainly include plate counting method, recombinant enzyme polymerase amplification method, real-time polymerase chain reaction method, enzyme-linked immunosorbent assay method, surface plasmon resonance method, quartz crys...
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IPC IPC(8): G01N33/569G01N33/558
CPCG01N33/56916G01N33/558
Inventor 吴正宗崔波袁超刘鹏飞于滨郭丽赵海波陶海腾
Owner QILU UNIV OF TECH