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Kit for detecting salmonella typhimurium and preparation method thereof

A technology for Salmonella and typhoid, which is applied to measuring devices, material analysis by observing the impact on chemical indicators, instruments, etc., can solve the problems of false positives, time-consuming, low sensitivity, etc.

Active Publication Date: 2020-07-03
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PCR technology can amplify the detection signal in a short time and has the characteristics of high sensitivity, but the operation process of PCR requires professionals and is prone to false positives
The ELISA method is based on the specific binding of antigens and antibodies for detection, without the need for large instruments, but it takes a long time and has low sensitivity
As an emerging detection technology, LAMP has the advantages of low cost, high sensitivity and specificity, but LAMP requires selective enrichment culture, so it is not suitable for on-site detection of samples

Method used

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  • Kit for detecting salmonella typhimurium and preparation method thereof
  • Kit for detecting salmonella typhimurium and preparation method thereof
  • Kit for detecting salmonella typhimurium and preparation method thereof

Examples

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preparation example Construction

[0036] The present invention also provides a preparation method of a kit for detecting Salmonella typhimurium, comprising:

[0037] Step 1: Fe 3 o 4 Preparation of nanoparticles

[0038] FeCl 3 ·6H 2 O, sodium citrate and CH 3 COONa·3H 2 O is magnetically stirred at room temperature in ethylene glycol until it is completely dissolved, and then the resulting homogeneous solution is transferred to a reaction kettle, preferably reacted in a 200°C oven for 10 hours; the dark brown solid matter is separated from the reaction solution with a permanent magnet, and the supernatant is used. Alternately wash with pure water and absolute ethanol for 3-5 times to obtain Fe with superparamagnetism 3 o 4 Nanoparticles; the FeCl 3 ·6H 2 O, sodium citrate and CH 3 COONa·3H 2 O mass ratio is preferably 2.16:0.5:3.34;

[0039] Step 2: Fe 3 o 4 Preparation of @Ag magnetic nanoparticles

[0040] Preparation of Fe by Liquid Phase Reduction 3 o 4@Ag nanoparticles, specifically: the...

Embodiment 1

[0053] Example 1 Salmonella apt-Fe 3 o 4 Preparation of @Ag NPs

[0054] First prepare the Fe 3 o 4 @Ag, 2.16g FeCl 3 ·6H 2 O, 0.5g sodium citrate and 3.34g CH 3 COONa·3H 2 O was magnetically stirred at room temperature in 40 mL of ethylene glycol until completely dissolved. Then transfer the obtained homogeneous solution to a reaction kettle and react in an oven at 200°C for 10 hours; use a permanent magnet to separate the dark brown solid matter from the reaction solution, wash it alternately with ultrapure water and absolute ethanol for 3-5 times, and obtain Superparamagnetic Fe 3 o 4 Nanoparticles.

[0055] Preparation of Fe by Liquid Phase Reduction 3 o 4 @Ag nanoparticles, 0.1g AgNO 3 and 2.5g polyvinylpyrrolidone k-30 were dissolved in 24mL polyethylene glycol, after fully dissolving, add 0.2g prepared Fe 3 o 4 Nanoparticles were dispersed in it, stirred at 100°C for 12h, the brown solid matter was separated from the reaction solution with a permanent mag...

Embodiment 2

[0057] The preparation of embodiment 2 urea / phenol red test paper:

[0058] Dissolve 0.1 g of phenol red and 3 g of urea in 60% ethanol solution. Soak neutral cotton filter paper in the above solution for 10 minutes at room temperature. The soaked test paper was laid flat and dried in an oven at 37°C. Store the prepared urea / phenol red test paper in a dry, sealed, cool and light-proof environment.

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Abstract

The invention provides a kit for detecting salmonella typhimurium and a preparation method of the kit, and belongs to the field of kits. The kit comprises an apt-Fe3O4@Ag NPs nanoprobe of salmonella,a hydrogen peroxide etching agent, a urease solution and urea / phenol red test paper. The invention also provides a preparation method of the kit for detecting salmonella typhimurium. The salmonella typhimurium is detected by using an immunomagnetic bead and nano particle composite structure, and color development is performed through test paper by using an enzyme inhibition method, so that the detection time is shortened, the coefficient of variation during quantitative detection is small, the lowest detection concentration is 21cfu.ML<-1>, the spiked recovery rate reaches 96.18%, the sensitivity is high, and the stability is good.

Description

technical field [0001] The invention belongs to the field of kits, in particular to a kit for detecting Salmonella typhimurium and a preparation method thereof. Background technique [0002] Salmonella typhimurium (S. typhimurium), a Gram-negative enteric bacterium, is one of the most common pathogens responsible for foodborne illness outbreaks. Salmonella usually contaminates food such as eggs, fruits and vegetables, meat and milk through animal feces. Salmonella infection can cause typical gastroenteritis reactions, such as abdominal pain, diarrhea, nausea, vomiting, fever, and other symptoms of gastroenteritis. According to reports, food poisoning caused by Salmonella is still on the rise. At present, this bacterium is a must-check pathogen for agricultural and sideline products such as eggs, meat and milk. [0003] At present, the traditional detection method of Salmonella in my country is mainly based on microbial culture method, which takes 3-5 days for the whole pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N21/78
CPCG01N21/78G01N33/54326G01N33/54346G01N33/56916G01N2021/7759G01N2333/255
Inventor 王娟赵超魏胜男
Owner JILIN UNIV
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