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Method for detecting salmonella typhimurium

A technology for Salmonella and Typhimurium, applied in the field of microbial detection, can solve the problems of high cost, low specificity and sensitivity, and achieve the effects of low detection limit, simple method and high specificity

Inactive Publication Date: 2017-07-21
UNIV OF JINAN
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AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of relatively low specificity and sensitivity and high cost in the method for detecting Salmonella typhimurium in the above prior art, the present invention provides a method for detecting Salmonella typhimurium with high specificity and sensitivity, low cost and fast detection speed. Methods

Method used

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Examples

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Embodiment 1

[0033] A method for detecting Salmonella typhimurium, specifically as follows:

[0034] a. Add sterilized water, 10× buffer (buffer), 1 μL primer chain and aptamer chain to the pre-prepared sterilized EP tube. Shake for 30s, incubate at 95°C for 5min, and slowly cool to room temperature;

[0035] b. Add target bacteria to the EP tube (concentrations are 9.8*10 5 cfu / mL, 9.8*10 4 cfu / mL, 9.8*10 3 cfu / mL, 9.8*10 2 cfu / mL, 9.8*10 1 cfu / mL), shaken for 30s, and incubated in a 37°C incubator for 2 h;

[0036] c. Continue to add HP (1 μL), helper (1 μL), endonuclease (1 μL), polymerase (1 μL) and dNTP (1 μL) to the EP tube; shake for 30 s, then mix the mixture The solution was further incubated in a 37°C incubator for 2 h;

[0037] d. Add heme, hydrogen peroxide and ABTS to the solution after the reaction in step c for 5 min;

[0038] Use a UV-vis spectrophotometer to detect the ABTS ultraviolet absorption signal at 410 nm, the detection range is 380nm-680nm, read the si...

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Abstract

The invention provides a method for detecting salmonella typhimurium. The method comprises the following steps: preparing an arch-shaped probe from aptamer and primer; then adding target bacteria into a reacted homogeneous solution, performing incubation at 37 DEG C for 2 hours, combining a target with the aptamer, releasing a primer sequence, opening HP, combining the HP onto a helper, and completing a subsequent EXPAR process, thus generating amplification of a signal; then adding heme, hydrogen peroxide and ABTS, and detecting ultraviolet absorption at 410 nm through a spectrometer. The detection method provided by the invention has the advantages of high detection speed, low detection limit, high specificity and the like.

Description

technical field [0001] The invention relates to a method for detecting Salmonella typhimurium, belonging to the technical field of microorganism detection. Background technique [0002] Salmonella is a Gram-negative bacterium with blunt ends, no spores, and generally no capsule. Except for Salmonella pullorum and Salmonella gallinarum typhi, others have flagella all over the body. Salmonella is a common zoonotic pathogen that can not only cause gastroenteritis, but also various syndromes such as sepsis, typhoid fever, and extraintestinal focal infection. In particular, pathogenic bacteria represented by Salmonella typhimurium have caused a huge threat in food safety, environmental monitoring, disease prevention, etc., and have caused great harm to human health, so they have received strong attention from people. [0003] At present, the methods mainly used to detect Salmonella typhimurium include biochemical identification and serotyping. However, this method has its o...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/10C12R1/42
CPCC12Q1/682C12Q1/689C12Q2525/205C12Q2521/307C12Q2531/119
Inventor 黄加栋裴倩倩刘素王玉王虹智郭玉娜
Owner UNIV OF JINAN
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