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Detection method capable of rapidly distinguishing LM (Listeria monocytogenes) from Listeria seeligeri

A technology of Listeria monocytogenes and a detection method, which is applied in the detection field of rapidly distinguishing Listeria monocytogenes from Listeria monocytogenes, can solve the problem of specific identification, indistinguishability, and difficulty in specific identification between species of Listeria monocytogenes. It can meet the problems of rapid identification, and achieve the effect of reducing the risk of human and environmental pollution, high degree of automation, and reducing the probability of exposure to food-borne pathogens.

Active Publication Date: 2017-02-22
INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The identification of Listeria monocytogenes mostly uses traditional biochemical reaction experiments and hemolysis tests. This method takes a long time, usually 5-7 days, and it is difficult to meet the needs of rapid identification.
In the national standard "GB 4789.30-2010", hemolysis test: Both Listeria monocytogenes and Listeria seeligeri (Listeria seeligeri) produce a narrow transparent hemolysis ring around the pricking point, and the two are difficult to distinguish; synergistic hemolysis Assay (cAMP): L. monocytogenes has increased hemolysis near the inoculum end of S. aureus and L. stutzeri has increased hemolysis, al

Method used

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  • Detection method capable of rapidly distinguishing LM (Listeria monocytogenes) from Listeria seeligeri
  • Detection method capable of rapidly distinguishing LM (Listeria monocytogenes) from Listeria seeligeri
  • Detection method capable of rapidly distinguishing LM (Listeria monocytogenes) from Listeria seeligeri

Examples

Experimental program
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Embodiment 1

[0023] (1) Culture preparation of Listeria monocytogenes ATCC 13932 and Listeria seeligeri ATCC 35967

[0024] Using an inoculation loop, aseptically transfer Listeria monocytogenes ATCC 13932 to 5 mL of sterilized nutrient broth medium, and culture on a shaker for 12 hours at 36°C and 150 r / min. Pipette all the culture into a 20mL Agilent crimp headspace sample bottle, seal it with a cap, and use it as the culture of Listeria monocytogenes to be tested.

[0025] Using an inoculation loop, transfer Listeria studii ATCC 35967 into 5 mL of sterilized nutrient broth medium by aseptic operation, and culture on a shaker at 36°C and 150 r / min for 12 hours. Pipette all the culture into a 20mL Agilent crimp-top headspace sample bottle, seal it with a cap, and use it as the culture of Listeria studii to be tested.

[0026] Among them, the nutritional broth components are: peptone 10.0g / L, beef powder 3.0g / L, sodium chloride 5.0g / L, glucose 1.0g / L. Purchased from Beijing Land Bridge T...

Embodiment 2

[0039] (1) Culture preparation of Listeria monocytogenes ATCC 19114 and Listeria seeligeri ATCC 35867

[0040] Using an inoculation loop, aseptically transfer Listeria monocytogenes ATCC 19114 to 5 mL of sterilized nutrient broth medium, and culture on a shaker for 12 hours at 36°C and 150 r / min. Pipette all the culture into a 20mL Agilent crimp headspace sample bottle, seal it with a cap, and use it as the culture of Listeria monocytogenes to be tested.

[0041] Using an inoculation loop, aseptically transfer Listeria studii ATCC 35867 to 5 mL of sterilized nutrient broth medium, and culture on a shaker for 12 hours at 36°C and 150 r / min. Pipette all the culture into a 20mL Agilent crimp-top headspace sample bottle, seal it with a cap, and use it as the culture of Listeria studii to be tested.

[0042] Nutrient Broth was purchased from Beijing Land Bridge Technology Co., Ltd.

[0043] (2) Headspace gas chromatography-mass spectrometry analysis of the culture

[0044] The A...

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Abstract

The invention discloses a detection method capable of rapidly distinguishing LM (Listeria monocytogenes) from Listeria seeligeri. According to the method, metabolic volatile products of the LM and the Listeria seeligeri obtained through liquid state fermentation are detected with an HS-GC-MS (headspace gas chromatographymass spectrometry) technology, and the LM is distinguished from the Listeria seeligeri according to different categories of the metabolic volatile products of the two bacteria. The identification time of the method is shortened by 1-2 days compared with the traditional biochemical experiments, the LM and the Listeria seeligeri on a plate can be distinguished rapidly, and the same-day screening is realized. The method is high in automation degree, effectively reduces the probability of contact of a frontline detector with food-sourced pathogenic bacteria and reduces the risks of pollution of the food-sourced pathogenic bacteria to humans and the environment.

Description

technical field [0001] The invention relates to a detection method for rapidly distinguishing Listeria monocytogenes and Listeria stutzeri by using headspace gas chromatography-mass spectrometry (HS-GC-MS) technology. Background technique [0002] Listeria monocytogenes (Listeria monocytogenes, LM), referred to as Listeria monocytogenes, is a foodborne pathogen that can cause zoonotic diseases. Listeria monocytogenes can contaminate many foods, including raw poultry, livestock meat, fermented sausages, and seafood. WHO listed it as one of the four major pathogenic bacteria in food in the 1990s. It is widely distributed in nature and can still grow and reproduce in an environment of 4°C. It is one of the main pathogenic bacteria that threaten human health in refrigerated food. It is very easy to contaminate food and cause food poisoning and listeriosis (caused by Listeria monocytogenes) The outbreaks of acute infectious diseases), especially in European and American countri...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 李正义贾俊涛崔淑华姜英辉赵丽青唐静程刚
Owner INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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