Method for rapidly detecting Salmonella in foods

A Salmonella, food technology, applied in the field of food testing, can solve the problems of time-consuming and laborious, and achieve the effect of low laboratory equipment requirements and low reagent cost

Inactive Publication Date: 2018-06-26
BEIJING UNIV OF TECH
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Problems solved by technology

Using traditional culture methods to detect Salmonella requires multi-step enrichment, separation, screening and biochemical ...

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  • Method for rapidly detecting Salmonella in foods
  • Method for rapidly detecting Salmonella in foods
  • Method for rapidly detecting Salmonella in foods

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Embodiment 1

[0026] 1 Materials and methods:

[0027] 1.1 Preparation of samples to be tested

[0028] 1.1.1 Bacterial culture

[0029] Salmonella enteritidiS (Salmonella enteritidiS) strain (50041) was purchased from China Culture Collection Center. Dip a little activated bacterial solution, streak on nutrient agar to separate the pure, pick a single colony to inoculate in TSB-YE liquid medium after culturing overnight at 37°C, and cultivate overnight at 37°C.

[0030] 1.1.2 Determination of initial addition amount

[0031] Take the cultured pure bacteria solution, and adjust the concentration of the bacteria solution to 0.5 McFarland with TSB-YE liquid medium. Use this concentration of bacterial liquid as the mother liquid, and continue to dilute with TSB-YE in a ratio of 10 times until the concentration of the bacterial liquid is 10 times that of the mother liquid. -8 . take 10 -5 、10 -6 100 μL of the dilution was inoculated on the surface of nutrient agar, two plates were inocul...

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Abstract

The invention provides a method for rapidly detecting Salmonella in foods and relates to the technical field of food detection. A PCR method is used to detect inVA genes of Salmonella in a to-be-tested food sample. The method includes the steps of adding the to-be-tested food sample into buffered peptone water to enrich the bacteria, then washing the bacteria with physiological saline, boiling lysed cells, and centrifuging to obtain the supernatant; taking the obtained supernatant as a PCR reaction template, and using detection primers of Salmonella inVA gene for PCR detection; wherein the detection primers are as follows: an upstream primer inVAF:5'-tcctccgctctgtctactta-3', and a downstream primer inVAR: 5'-accgaaatattcattgacgtt-3'. The detection results of the method are consistent withthose of a conventional culture method and a BAX system, the detection limit is 100cfu/25g, and the accuracy is 100%.

Description

technical field [0001] The invention relates to the technical field of food detection, in particular to a method for rapidly detecting salmonella in food. Background technique [0002] Salmonella is one of the main foodborne pathogenic microorganisms, which can easily contaminate meat, fish, poultry, milk, and egg foods. The time limit requirements for import and export food testing are very strong, otherwise it will affect the performance of the contract or reduce the value of the goods. Especially for fresh food, such as fresh aquatic products, chilled chicken, etc., it is even more required to shorten the detection time. Using traditional culture methods to detect Salmonella requires multi-step enrichment, separation, screening and biochemical identification, and if necessary, serological identification, which usually takes 4 to 6 days, which is time-consuming and laborious. The PCR method developed in recent years can greatly shorten the detection time, and has high sp...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04C12R1/42
CPCC12Q1/686C12Q1/689
Inventor 孙永军王湛侯磊朱可欣杜心培
Owner BEIJING UNIV OF TECH
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