A rapid detection method for distinguishing Salmonella typhimurium from Proteus mirabilis
A technology of Proteus mirabilis and Salmonella, applied in measuring devices, material separation, and analysis of materials, can solve problems affecting PCR amplification results, achieve high automation, reduce the risk of human and environmental pollution, and quickly screen Effect
Active Publication Date: 2019-02-22
INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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Problems solved by technology
PCR technology is a fast, sensitive, and specific technology, but at present, this technology still relies on the pre-enrichment step of the traditional method. The enrichment solution often contains PCR inhibitors, which affects the PCR amplification results.
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Embodiment 1
[0024] (1) Culture preparation of Salmonella typhimurium ATCC 25241 and Proteus mirabilis ATCC 25933
[0025] The aseptic operation of the inoculation loop was used to transfer Salmonella typhimurium ATCC 25241 into 5 mL of sterilized nutrient broth, and cultured on a shaking table at 36°C and 120 r / min for 12 hours. Draw all the culture and transfer it into a 20mL Agilent crimp headspace sample bottle, seal it with a cap, and use it as the culture of Salmonella typhimurium to be tested.
[0026] Transfer Proteus mirabilis ATCC 25933 into 5 mL of sterilized nutrient broth by aseptic operation of the inoculation loop, and culture on a shaker at 36°C and 120 r / min for 12 hours. Pipette all the culture into a 20mL Agilent crimp headspace sample bottle, seal it with a cap, and use it as the culture of Proteus mirabilis to be tested.
[0027] Among them, the nutrient broth components are: peptone 10.0g / L, beef powder 3.0g / L, sodium chloride 5.0g / L, glucose 1.0g / L, purchased from B...
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The invention discloses a detection method for rapidly distinguishing salmonella typhimurium and proteus mirabilis. The detection method comprises the following steps: utilizing a headspace gas chromatography-mass spectrometry linked technique to detect volatile products produced from liquid-state fermentation of the salmonella typhimurium and the proteus mirabilis, and distinguishing the salmonella typhimurium and the proteus mirabilis according to the difference of the volatile products produced by two pathogenic bacteria. The detection method disclosed by the invention has the advantages that the identification time is shortened by 1-2 days compared with that of the existing traditional biochemical experiment, the salmonella typhimurium and the proteus mirabilis on a plate can be rapidly distinguished and fast screening is realized; the automation degree is high, the probability of contact of first-line detection personnel with the pathogenic bacteria is effectively reduced and the risk of the food-borne pathogenic bacteria to personnel and environmental pollution is reduced.
Description
technical field [0001] The invention relates to a detection method for quickly distinguishing Salmonella typhimurium and Proteus mirabilis by using headspace gas chromatography-mass spectrometry (HS-GC-MS) technology. Background technique [0002] Salmonella (Salmonella) is one of the main intestinal pathogens, which can easily cause food poisoning. In my country, food poisoning caused by Salmonella occupies the first place in bacterial food poisoning. At present, more than 2,000 serotypes of Salmonella have been isolated in the world, among which Salmonella typhimurium and Salmonella enteritidis are the most common. Salmonella typhimurium, a group of non-adaptive or pantropic Salmonella, has a wide range of hosts and is currently one of the highest isolation rates in the world. The bacterium can cause various infectious diseases of poultry and mammals, and can also cause human infection, which has important public health significance. The detection of Salmonella typhimur...
Claims
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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 贾俊涛赵丽青李正义雷质文唐静马云
Owner INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU