Cold water gel test paper sheet for rapid detection of Listeria monocytogenes in food and tableware and its preparation method and application

A technology of Listeria monocytogenes and cold water gel is applied in the field of cold water gel test strips and preparation thereof, which can solve the problem that the detection of Listeria monocytogenes on the surface of tableware cannot be used, the quantitative detection requirements cannot be met, and only qualitative detection can be performed. Detection and other problems, to achieve the effect of low cost, strong specificity and high sensitivity

Active Publication Date: 2020-04-10
HUBEI UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above method requires special detection equipment, which is not convenient for daily on-site rapid detection requirements.
The patent application with the application number 201510274726.9 discloses a "test paper for rapid detection of Listeria monocytogenes and its preparation and application". In the functional area, a fluid channel is arranged between two adjacent areas, and a substrate layer containing the substrate 5-bromo-4-chloro-3-indoxylinositol ammonium phosphate X-InP is arranged in the boundary of the substrate area, but The detection test paper disclosed in this prior art is mainly used for the qualitative detection of Listeria monocytogenes contamination in drinking water, meat products, dairy products, and vegetables, and cannot be used for the detection of Listeria monocytogenes on the surface of tableware, and the detection Test strips can only be used for qualitative testing and cannot meet the needs of quantitative testing

Method used

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  • Cold water gel test paper sheet for rapid detection of Listeria monocytogenes in food and tableware and its preparation method and application
  • Cold water gel test paper sheet for rapid detection of Listeria monocytogenes in food and tableware and its preparation method and application
  • Cold water gel test paper sheet for rapid detection of Listeria monocytogenes in food and tableware and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Chromogenic agent (2R,3S,4R,5R,6R)-1,2,3,4,5-pentahydroxycyclohexyl-(2-nitro-4-fluorophenyl)phosphate is prepared as follows , including the following steps:

[0047] (1) Dissolve 1 mmol of reactant A (1,2,3,4,5,6-hexamethoxymethylene-inositol) in 2.5 mmol of anhydrous diisopropylamine and a small amount of anhydrous chloroform to obtain the reactant System A, and then freeze the reactant system A at -70°C;

[0048] (2) At a low temperature of -70°C, 1.25 mmol of reactant B (methyl dichlorophosphite) was added dropwise to the reactant A system in an anhydrous nitrogen atmosphere, vigorously stirred, and reacted for 6 hours;

[0049] (3) Place the system after the reaction in step (2) at room temperature. After the reaction system reaches room temperature, continue to add 2 mmol of reactant C (2-nitro-4-fluorophenol) dissolved in anhydrous Chloroform solution, continue to stir and react at room temperature for 12 hours to generate phosphite substances;

[0050] (4) Ad...

Embodiment 2

[0054] A kind of preparation method of listeria monocytogenes rapid detection cold water gel test paper sheet of the present embodiment, described method comprises the following steps:

[0055] (1) Preparation of carrier paper sheets: Cut the chromatographic filter paper into a size of 5 cm × 5 cm, irradiate or autoclave (after high pressure, dry at a constant temperature of 45° C. in vacuum) for later use;

[0056] (2) Preparation of the inner packaging bag: press the high-temperature-resistant polypropylene plastic film bag into a joint bag with a size of 5.1cm×5.1cm according to the test requirements;

[0057] (3) Preparation of basal medium (1L): Each liter of medium contains tryptone 5.0-6.0g, beef powder 5.0-6.0g, polyvalent peptone 3.0-5.0g, sodium chloride 8.0-10.0g, dihydrogen phosphate Potassium 1.0-1.5g, disodium hydrogen phosphate 5.0-7.0g, glycine 8.0-10.0g, lithium chloride 1.0-2.0g, phenylethyl alcohol 2.0-3.0g, escin 1.0-1.5g;

[0058] (4) The chromogen (2R, 3...

Embodiment 3

[0064] Embodiment 3 Rapidly detects the interfering test of test paper sheet

[0065] At the same time, shake flasks of Escherichia coli, Staphylococcus aureus, Salmonella, Vibrio parahaemolyticus, and Listeria monocytogenes were cultivated overnight, and 0.5mL of the bacterial suspension was centrifuged at 8000r / min for 5min, and diluted tenfold to A certain multiple (10 2 cfu / mL), followed by mixing 0.5 mL of Escherichia coli, Staphylococcus aureus, Salmonella, and Vibrio parahaemolyticus suspension with 0.5 mL of Listeria monocytogenes suspension, and resuspended in 1 mL of sterile water , drop 1mL of the bacterial suspension onto the detection paper, incubate at 36°C±1°C for 24-36h, observe the growth of the colonies on the paper, and study whether other bacteria interfere with the growth of Listeria monocytogenes, set up 5 parallel group, the test was repeated 3 times, and the test results showed that as long as there was Listeria monocytogenes in the mixed bacterial sol...

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Abstract

The invention relates to a cold hydrogel test strip for rapid detection of Listeria monocytogenes in food and tableware, and a preparation method and application thereof, belonging to the technical field of microorganism safety examination and detection. According to the invention, cold hydrogel is utilized, chromatographic filter paper is used as a carrier, and components of a medium, a developer and the cold hydrogel are mixed and are then uniformly loaded on the chromatographic filter paper; a sample can be uniformly distributed in the cold hydrogel, and distinct bacterium colony characteristics are formed on the surface of the cold hydrogel; thus, rapid, accurate, qualitative and quantitative determination of Listeria monocytogenes is realized. A novel compound (2R,3S,4R,5R,6R)-1,2,3,4,5-pentahydroxycyclohexyl-(2-nitro-4-fluorophenyl)phosphate is used as the developer, so the test strip provided by the invention has substantially superior reactive sensitivity and specificity to those of a traditional chromogenic medium for Listeria monocytogenes, resists irradiation and has better chemical stability. Moreover, the test strip prepared in the invention has low cost; and the preparation method is simple and is completed in one step, and detection time only needs 24 to 36 h.

Description

technical field [0001] The invention belongs to the technical field of microbial safety inspection and detection, and in particular relates to a rapid detection paper sheet, more specifically, the invention relates to a qualitative and quantitative detection device specially used for the rapid detection of Listeria monocytogenes in food and tableware Cold water gel test paper sheet and its preparation method and application. Background technique [0002] Listeria monocytogenes is widely distributed, and Listeria monocytogenes is the only one capable of causing disease in humans. Listeria monocytogenes is a common soil bacterium in which it is a saprophyte that feeds on dead and decaying organic matter. It is also a contaminant in certain foods (mainly fresh milk products) and can cause severe food poisoning. Listeria monocytogenes is a zoonotic pathogen. It can cause listeriosis in humans and animals, and the main manifestations after infection are sepsis, meningitis and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12R1/01
Inventor 张毅代俊周梦舟唐景峰朱莹张超
Owner HUBEI UNIV OF TECH
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