Method for checking monocyte hyperplasia Listeria

A technology of mononucleosis and living bacteria, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc.

Inactive Publication Date: 2008-09-10
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention provides a method for detecting viable Listeria monocytogenes in order to solve the problem that existing methods for detecting viable Listeria monocytogenes are prone to false positives

Method used

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  • Method for checking monocyte hyperplasia Listeria
  • Method for checking monocyte hyperplasia Listeria
  • Method for checking monocyte hyperplasia Listeria

Examples

Experimental program
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specific Embodiment approach 1

[0020] Specific embodiment one: the method for the detection of Listeria monocytogenes viable bacteria in this embodiment is carried out according to the following steps:

[0021] 1. Take 25g of the article to be tested and place it in 225mL tryptone soybean yeast extract broth for 8 hours, then take 4mL of the culture solution and put it in a 5mL epoxy resin tube, and centrifuge at room temperature at a speed of 14000r / min 3min, get the fungus slime;

[0022] 2. Resuspend the obtained bacteria slime in a mixture of 3 mL of methanol, ether and ammonia, then resuspend in 1 mL of lysate, then add 0.2 g of glass beads with a diameter of 0.2 mm, and vortex at 2000 rpm 5min. Then centrifuge for 3min with the relative centrifugal force of 14000r / min;

[0023] 3. Take 700 μL of the suspension after centrifugation and mix it with the same volume of water-saturated phenol with a pH value of 5.5 in a 1.5 mL epoxy resin tube, incubate at 68°C for 5 minutes, and then centrifuge at room ...

specific Embodiment approach 2

[0035] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the DEPC water in step 7 is MiliQ pure water treated with DEPC (diethypyrocarbonate, diethyl pyrocarbonate) and sterilized by high temperature and high pressure. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0036] Embodiment 3: The difference between this embodiment and Embodiment 1 is: the volume ratio of methanol:ether:ammonia in the mixture of methanol, ether and ammonia in step 2 is 1:1:1. Other steps and parameters are the same as those in Embodiment 1.

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Abstract

The invention discloses a method for detecting viable Listeria monocytogene, relating to a method for detecting Listeria. The invention solves the problem that the prior method for detecting viable Listeria monocytogenes is easy to produce false positive. The method for detecting viable Listeria monocytogenes is carried out as following steps: after extracting bacterial sludge from articles to be detected, extracting RNA to implement retrotransposon and real time PCR; accordingly, a collection of illustrative plates which are capable of determining whether the articles contain viable Listeria monocytogenes or not. The method has the advantages of high sensitivity, good specificity, simple operation, and comparatively short period. The method meets the requirements of pathogen rapid detection in food industry.

Description

technical field [0001] The invention relates to a method for detecting Listeria. Background technique [0002] Listeria monocytogenes (Listeria monocytogenes, Lm), referred to as Listeria monocytogenes, is a facultative anaerobic Gram-positive bacteria, its pH value growth range is 4.39 ~ 9.40, in It can also grow at 4°C, thus increasing the hazard to refrigerated food. Listeria monocytogenes widely exists in soil, animals, and aquatic products, and is mainly transmitted through foods such as milk and dairy products, vegetables, aquatic products, and meat products. Because of its unique physiological characteristics, it can cause human meningitis, septicemia and other diseases, especially pregnant women, newborns and patients with low immune function are more susceptible to infection, with a fatality rate of more than 30%. [0003] The existing method for detecting Listeria monocytogenes requires at least 4 to 7 days for the whole process, the detection limit is not sensit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12Q1/68
Inventor 姜毓君闫冰韩希妍曲妍妍王明娜相丽吕琦毕宇涵
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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