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A co-enrichment medium ses

A technology of enrichment medium and basic medium, which is applied in the field of agricultural product testing, can solve the problems that high-throughput detection methods are difficult to detect and cannot bypass the enrichment, so as to save quantity, avoid the interference of bacteria, and test samples wide range of effects

Active Publication Date: 2019-09-27
SHANGHAI ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, enrichment culture is an essential step in microbial detection. Existing new technologies for rapid detection of various microorganisms, such as various polymerase chain reaction (PCR), gene chips, enzyme-linked immunosorbent (ELISA) and other methods Due to the limitation of self-detection ability (detection limit 10 2 ~10 4 ) and the interference of background microorganisms cannot bypass the enrichment process of enrichment
Moreover, for different pathogenic bacteria to be tested, the enrichment solution and selective medium used in the sample are also different. This kind of enrichment solution that only enriches for a specific bacterial species largely limits high-throughput detection. The application of technologies (such as multiplex PCR and gene chip detection technology) has made it difficult for high-throughput detection methods to be used in the detection of actual samples

Method used

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  • A co-enrichment medium ses
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  • A co-enrichment medium ses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: SES monoenrichment effect identification

[0036] Preparation of SES medium: (1) Basal medium: 20 parts of tryptone, 1 part of glucose, 5 parts of NaCl, 4 parts of dipotassium hydrogen phosphate, 1.5 parts of potassium dihydrogen phosphate; (2) Inhibitor: No. 3 bile salt 1.2 parts, 20 parts of novobiocin sodium salt, 0.5 parts of lithium chloride, 0.1 parts of potassium tellurite; (3) distilled water, 1000 parts. Weigh tryptone, glucose, NaCl, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, No. 3 bile salt, lithium chloride, add water and stir evenly, autoclave at 121°C, cool to room temperature, in the above mixture, no Add novobiocin sodium salt and potassium tellurite to the bacteria, and mix well to prepare the required co-enrichment medium (SES).

[0037] SES monoenrichment culture: take the overnight culture of Salmonella enteritidis (RV), Escherichia coli (mEC) O157:H7 and Shigella flexneri (GN) and inoculate 100ml of SES and their res...

Embodiment 2

[0039] Example 2: Identification of SES compound bacterial enrichment effect

[0040] Preparation of SES medium: refer to Example 1.

[0041]SES compound enrichment culture: Three target bacteria, Salmonella enteritidis, Escherichia coli O157:H7 and Shigella flexneri, were inserted into SES at different concentration ratios. The proportions of the compound inoculation of the above three kinds of bacteria were as follows: 1:1:1, 1:10:100, 10:100:1 and 100:1:10. Cultivate at 36°C and 180rpm / min for 24h, and take samples every 4h for colony counting. The enumeration medium of each target bacteria was selected from its own selective plate: BS for Salmonella enteritidis, CT-SMAC for Escherichia coli O157:H7, and selective plate for Shigella flexneri.

[0042] SES compound bacterial enrichment effect identification: according to the colony counting results of each time period, the growth curve was drawn ( figure 2 ), comparing the growth ability of target bacteria with different ...

Embodiment 3

[0043] Example 3: Detection of 3 foodborne pathogens in frozen pork by SES combined with multiplex PCR

[0044] Preparation of SES medium: refer to Example 1.

[0045] Treatment of samples to be tested: Take 10 g of commercially available raw pork, soak the pork in alcohol, disinfect and air-dry, inoculate Salmonella Enteritidis, Escherichia coli O157:H7, and Shigella flexneri respectively at a final concentration of about 10 CFU / g, and air-dry to The bacterial solution was completely absorbed, and stored in a homogeneous bag at -20°C for 7 days.

[0046] SES enrichment culture and effect identification: put 10g of the artificially polluted pork sample in 90ml SES, and culture it at 36°C for 24h. From 12h, take the enrichment solution every 4h to count the viable bacteria of the above three target bacteria, and identify the SES. Bacteria effect ( image 3 ). The three target bacteria can still be enriched by SES after being frozen in pork at -20°C for 7 days, and the final ...

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Abstract

The invention provides a coenrichment bacterial culture medium SES, which consists of a basic culture medium, an inhibitor and water. The coenrichment bacterial culture medium SES can simultaneously enrich three bacteria including salmonella enteritidis, escherichia coli O157:H7 and Shigella so as to realize a purpose of simultaneously detecting various pathogenic bacteria.

Description

technical field [0001] The invention relates to the field of agricultural product detection, in particular to a co-enrichment culture medium SES capable of simultaneously enriching Salmonella enteritidis, Escherichia coli O157:H7 and Shigella flexneri. Background technique [0002] Salmonella (Salmonella, SM), Escherichia coli (EC) and Shigella (Shigella, SH) are common foodborne pathogens. Food poisoning incidents caused by these pathogenic bacteria have occurred many times at home and abroad. For example, in 2006, 199 people fell ill and 3 died in 26 states of the United States because of eating spinach contaminated by E. coli O157:H7. In 2007, 230 people became ill in Europe and Australia due to eating carrots contaminated with Shigella; in 2012, 261 people became ill and 3 died in several states of the United States due to eating cantaloupe contaminated with Salmonella. These three pathogenic bacteria are widely distributed in nature, and the risk of food contamination ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12Q1/10C12R1/42C12R1/19C12R1/01
CPCC12Q1/10Y02A50/30
Inventor 索玉娟高士刚周昌艳瞿洋白冰
Owner SHANGHAI ACAD OF AGRI SCI
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