KR101996906B1 - Selective Enrichment Media for Listeria monocytogenes - Google Patents

Abstract

The present invention relates to a medium for selectively enriching Listeria monocytogenes. Using the medium according to the present invention, only Listeria monocytogenes, which is the only pathogenic microorganism having infectivity against humans, can be selectively enriched among Listeria species, so that Listeria monocytogenes can be effectively used in the pre-enrichment step before detection.

Description

Listeria monocytogenes selective enrichment medium {Selective Enrichment Media for Listeria monocytogenes}

The present invention relates to a Listeria monocytogenes selective enrichment medium for promoting only the growth of Listeria monocytogenes among the species belonging to the genus Listeria and for maintaining or slowing the growth of other species belonging to the genus Listeria.

Listeria fungi include L. monocytogenes, L. ivanovii , L. innocua, L. seeligeri , L. grayi , Listeria monocytogenes , , and 6 species of Listeria Welsh Mary (L. welshimeri), L. Ivanova ratio (L. ivanovii) is a non-Listeria spp Ivano Ivano ratio (L. ivanovii subs. ivanovii ) and Listeria ivanoviasi L. ivanovii subs. londoniensis ) are divided into two subspecies.

Listeria (Listeria) is widely distributed in natural environment and foods. In healthy adults with normal immune function, severe infection such as septicemia and meningitis occurs in patients whose infection is not caused, and cellular immunity is decreased mainly. Or through infection with newborn infants. And the infection is caused by a mainly contaminated food, Listeria, only L. monocytogenes jeneseu (Listeria monocytogenes) and L. Ivanova ratio (L. ivanovii) Species of the pathogen and of the L. monocytogenes jeneseu (Listeria monocytogenes ) are known to cause infection in humans.

Listeria monocytogenes is a Gram-positive intracellular parasitic bacterium that is rod-shaped, withstands freezing and drying with cold bacteria growing at 1 to 44 ° C, and can withstand relatively long periods of dryness. Listeria monocytogenes causes infectious food poisoning. In 1983, Listeria, which was found to be infectible to humans by food, became one of the major diseases in public health. Listeria monocytogenes, the causative agent of infection, was first identified in 1920. This bacterium causes no symptoms or mild fever, abdominal pain, diarrhea, vomiting, and headache in healthy people. However, special attention should be paid to pregnant women, newborns, the elderly and patients with impaired immune function. The germs have recently been attracted to the public by massive food poisoning incidents in the United States and the risk of foodborne illness due to the germs has become greater as the consumption of simple cooked food to eat without further cooking increases. Listeria monocytogenes is a bacterium that causes bacterial meningitis, and is the most feared with a mortality rate of 22%. In addition, it is one of the three major causative agents of neonatal meningitis, the second most common bacterial infection in meningitis occurring in patients over 50 years of age, and the most common bacterial meningitis that occurs in immunocompromised patients with lymphoma, organ transplantation, to be. Other brain meningitis, brain abscess, endocarditis, and local infection may occur. Recent studies suggest that acute gastroenteritis may occur in a population, leading to acute diarrhea in normal people. Therefore, the detection of Listeria monocytogenes is important in terms of food hygiene and public health.

Currently, the detection standard of Listeria monocytogenes in food is based on non-detection in 25 g food samples in Korea, USA and Australia. Currently, the internationally recognized Listeria monocytogenes The detection method is the US Department of Agriculture-Food Safety and Inspection Service (USDA-FSIS) and the Bacteriological Analytical Manual (BAM) method developed by the FDA. In Korea, the foodstuff, Listeria monocytogenes The detection method is known, as with USDA-FSIS and BAM, the pre-coagulation time is 48 hours. However, Listeria monocytogenes has a characteristic of slowly growing, and the growth of bacteria by microorganism guns in food affects the growth of non-pathogenic Listeria species ( Listeria spp., Which grows faster than the hospital Listeria monocytogenes) It has been reported that the negative results can be obtained when the growth of the microorganisms is disturbed by the presence of a small amount of pathogenic bacteria.

Accordingly, a problem to be solved by the present invention is to provide a medium for selectively enriching Listeria monocytogenes only in Listeria monocytogenes pre-fermentation step so that no false negative result is obtained in the detection of Listeria monocytogenes.

In order to solve the above problems, the present invention provides a medium for selectively enriching Listeria monocytogenes.

More specifically, the present invention relates to a Listeria monocytogenes enrichment medium containing a carbon source, a nitrogen source and a phosphorus source,

Comprising sucrose as said carbon source;

Wherein the nitrogen source comprises a beef extract; And

Wherein the phosphorus contains potassium dihydrogen phosphate,

Lt; RTI ID = 0.0 > Listeria monocytogenes < / RTI >

Listeria monocytogenes selective enrichment medium.

The medium according to the present invention selectively enriches Listeria monocytogenes as a Listeria monocytogenes selective enrichment medium.

In the present invention, the term " selective enrichment medium " is a medium for enhancing the proliferation of a specific species when one or more microorganisms are mixed. In some cases, it is a medium which does not affect the proliferation of other species mixed with a certain extra-species, or inhibits the proliferation.

According to the present invention, the specific species is Listeria monocytogenes. Other species that are mixed out of a particular species may include Lyserian monocytogenes and similar species that are similar in propagation conditions (eg, temperature, humidity, nutrient source, etc.), and may contain species that compete with Listeria monocytogenes for replication, A species that causes false negativity when detecting cytogenes may be included. For example, Escherichia coli (e.g., ATCC 25922), Salmonella enteritidis (e.g., ATCC 13076), Shigella sonnei) (eg. ATCC 9290), Vibrio para H. Molina di kusu (Vibrio parahaemolyticus) (eg. ATCC 17803), Enterococcus coli yeosi California urticae (Yersinia enterocolitica) (eg. ATCC 9610), Escherichia coli 0157 (Escherichia E. coli O157) (e.g., ATCC 43894), Enterobacter aerogenes (e.g., ATCC 13048), Citrobacter freundii ) (eg, ATCC 8090); Or Gram positive bacteria including Bacillus cereus (eg ATCC 11778), Staphylococcus aureus (eg ATCC 12600), L. innocua (eg ATCC 33090) And the like, preferably Listeria inocuas.

The Listeria monocytogenes selective enrichment medium according to the present invention comprises sucrose as a carbon source; A beef extract as a nitrogen source; And potassium dihydrogen phosphate as a source.

The inventors have included three components, sucrose as a carbon source; A beef extract as a nitrogen source; And when potassium dihydrogen phosphate was included as a source, it was confirmed that there was an excellent effect as a medium for selectively enriching Listeria monocytogenes.

The sucrose is a polymerization molecule of glucose and fructose. The sucrose may be produced by a method known in the art or may be commercially available products such as ham, sugar, syrup, honey, caramel, etc. May be used. The sucrose is preferably contained at a concentration of 0.2 to 0.5% (g / L, mass of the ingredient relative to the total volume of the medium). If the sucrose is less than 0.2%, it is difficult to exhibit a significant antimicrobial effect .

The beef extract is obtained by extracting a water-soluble extract from beef (for example, a muscle portion), and may be manufactured by a method known in the art or may be a commercially available product. A variety of formulations may be used, including, for example, concentrates, powders, liquids, and the like, but are not limited thereto. The beef extract is preferably contained at a concentration of 0.6 - 1.8% (g / L, mass of the ingredient relative to the total volume of the medium). If the beef extract is less than 0.6%, it is difficult to exhibit a significant antimicrobial effect .

The potassium dihydrogen phosphate includes compounds of the formula KH ₂ PO ₄ or H ₂ KO ₄ P, salts thereof, hydrates thereof, and derivatives thereof. It may be produced by methods known in the art or may be commercially available products. It is preferable that potassium dihydrogen phosphate is contained at a concentration of 0.25 - 0.5% (g / L, mass of the component relative to the total volume of the medium) concentration. If it is less than 0.25%, it is difficult to exhibit a significant antimicrobial effect. It is economical.

Additionally, the Listeria monocytogenes selective enrichment medium according to the present invention comprises sodium-containing salts, more preferably sodium chloride.

The sodium chloride includes compounds of the formula Nacl, hydrates thereof, and derivatives thereof. It may be produced by methods known in the art or may be commercially available products.

Preferably, the concentration of sodium chloride is included in a concentration of 0.5 - 3% (g / L, mass of the component relative to the total volume of the medium). If the concentration is less than 0.5%, the effect is insufficient. to be.

In addition to the above-mentioned components, the following components may be further included as long as the object of the present invention is not impaired.

The carbon source is a component mainly used as an energy source and a constituent element of cells for growth or proliferation. Examples of the carbon source include galactose, xylose, maltose, glycerol, crude glycerol (biodiesel waste Crude glycerol), and the like. However, it is preferable that it does not substantially contain glucose, lactose and / or fructose. By "substantially free" is meant a concentration of less than or equal to 2%, more preferably less than or equal to 1%, more preferably less than or equal to 0.5%, most preferably less than or equal to 0% .

The nitrogen source may be an inorganic nitrogen source, an organic nitrogen source, or a combination thereof, which is necessary for biosynthesis of the protein. Examples of the nitrogen source include a corn steep liquor, a malt extract, a peptone, But are not limited to, organic nitrogen sources such as ammonia, ammonium nitrate, ammonium nitrate, ammonium nitrate, sodium nitrate, and Urea. However, it is preferable that substantially no yeast extract is contained. By "substantially free" is meant a concentration of 5% or less, more preferably 3% or less, even more preferably 1% or less, most preferably 0% (ie, no inclusion) .

The number may include any source that can supply phosphorus (P), and may act to minimize the pH change of the medium using the buffering action in the neutral region. However, it is preferable that it does not substantially contain di-potassium hydrogen phosphate. By "substantially free" is meant a concentration of less than or equal to 2%, more preferably less than or equal to 1%, more preferably less than or equal to 0.5%, most preferably less than or equal to 0% .

In addition, metal salts, vitamins and precursors thereof, and combinations thereof may be further included. The metal salt may include, for example, magnesium sulfate or iron sulfate, but is not limited thereto.

In addition, antibiotics may additionally be included, including, but not limited to, cycloheximide.

Most preferably, the Listeria monocytogenes selective enrichment medium according to the invention is selected from the group consisting of tryptone peptone, peptone, sucrose, potassium dihydrogen phosphate, beef extract, acriflavine hydrochloride, cyclohexamide, Chloride.

More specifically, 10-20 g / L of tryptone peptone, 1-5 g / L of peptone, 1-5 g / L of sucrose, 1-10 g / L of potassium dihydrogen phosphate, 5-20 g of beef extract / L, acroleplavin hydrochloric acid 0.001 - 0.05 g / L, cyclohexamide 0.01 - 1 g / L, naringic acid 0.01 - 1 g / L and sodium chloride 20 - 50 g / L.

Can be effectively used for isolating, identifying or detecting Listeria monocytogenes by proliferating or growing Listeria monocytogenes in a Listeria monocytogenes selective enrichment medium according to the present invention.

More specifically, by conducting a step of culturing a sample suspected of containing Listeria monocytogenes in a Listeria monocytogenes selective enrichment medium, the presence of Listeria monocytogenes in the sample can be confirmed in a short time with high efficiency and no false-negative.

The term 'sample' is included without limitation as long as it is a subject that is expected to include Listeria monocytogenes, for example, various types of foods in which Listeria monocytogenes contamination is expected.

The term 'separation' is a step for more selectively separating the enriched Listeria monocytogenes in the sample. Various methods known in the art may be used. For example, a selective medium may be used. As the selective medium, For example, LPM agar, Oxford agar, or PALCAM agar. After incubation at 35 ± 1 ° C. (or 30 ° C.) for 48 ± 2 hours, a colony surrounded by dark brown or black circles Can be used.

The term 'identification', 'detection' or 'confirmation' is used to identify the presence of Listeria monocytogenes in the sample, and various methods known in the art may be used. For example, Gram stain, Catalase test , β-hemolytic test, CAMP test, motility test, Mannitol resolution, Rhamnose resolution, Xylose resolution, Vitek analysis, O serum aggregation reaction, or H serum aggregation reaction But are not limited thereto.

More specifically, the present invention relates to a method for producing Listeria monocytogenes by pre-incubating Listeria monocytogenes; And

And detecting Listeria monocytogenes.

A method for detecting Listeria monocytogenes can be provided.

Between the pre-enriching step and the detecting step, the above-described separating step may be further included.

In addition, the present invention can provide a kit comprising Listeria monocytogenes selective enrichment medium.

The kit may be a kit for Listeria monocytogenes selective assimilation or a kit for detection, identification or confirmation of Listeria monocytogenesis.

The kit may include instructions describing a culture method, culture conditions, and the like in which Listeria monocytogenes selective enrichment medium can be applied to cultivate Listeria monocytogenes properly.

Using the culture medium according to the present invention, only Listeria monocytogenes is selectively and efficiently enriched, and thus Listeria monocytogenes, which is the only Listeria species, can be selectively detected.

BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments of the invention and, together with the description of the invention, It should not be construed as limited.
1 is a graph showing changes in the amount of Listeria monocytogenes detected in a mixed culture of four species of interstitial Listeria.
●: LM (Listeria monocytogenes), ×: LM (Listeria monocytogenes) + LN (Listeria monocytogenes) mix, ▲: LM (Listeria monocytogenes) LV (Listeria monocytogenes) + LN (Listeria monocytogenes) + LS (Listeria monocytogenes) + LV (Listeria monocytogenes) + LV (Listeria monocytogenes)
Figure 2 is a graph showing the relationship between the concentration of sucrose (a-1), sodium chloride concentration (b), nitrogen source component (c), beef extract concentration (c- (LM) and Listeria monocytogenes (LN) according to the present invention.

Hereinafter, specific examples of examples and experimental examples will be described in order to explain the present invention in detail. However, the embodiments or experiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the above-mentioned examples. It is to be understood that both the foregoing description and the following examples are provided for a more complete understanding of the present invention to those skilled in the art. Accordingly, various equivalents and variations Examples should be understood.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.

Example  One. Listeria  Four species of inter species competition analysis

Listeria monocytogenes, LN (listeria licorice), LS (listeria siligeri), and LV (listeria ivanovi) were purchased from National Type Culture Collection (ATCC, Manassas, Va., USA) ).

Listeria monocytogenes and Listeria monocytogenes. The growth of each strain was measured by ABI 7500 Real time PCR (Thermo Fisher Scientific, Bremen, Germany) at 0 h, 10 h, 18 h, 21 h, 24 h, 27 h, 30 h, Germany). The statistical significance was verified by ANOVA Duncan test using SPSS ( P <0.05).

The results are shown in Fig.

When Listeria monocytogenes (LM) and Listeria inocua (LN) were mixed at a ratio of 1: 1, Listeria monocytogenes (LM), which is the most rapidly growing among the four listeria species, ), And these results were prominent from 21 hours after the growth (FIG. 1 (a)).

Even when Listeria monocytogenes (LM), Listeria monocytogenes (LN), Listeria siligeri (LS) and Listeria ivanovi (LV) were mixed together in a ratio of 1: 1: 1: 1, From the passage of time, the growth of L. monocytogenes (LM) began to slow (Fig. 1 (d)).

On the other hand, in a 1: 1 mixed culture of Listeria monocytogenes (LM) and Listeria siligeri (LS), and a 1: 1 mixed culture of Listeria monocytogenes (LM) and Listeria ivanovi (LV) (Fig. 1 (b) and Fig. 1 (c)).

 Therefore, in order to increase only the detection rate of the intended Listeria monocytogenes in consideration of the competition among the four interspecies of Listeria, it was confirmed that a medium that supports the growth of Listeria monocytogenes while suppressing the growth of Listeria inocua (LN) is required .

Example  2. Listeria Monosaitogenes  Selection Enrichment  Badge development

By varying the carbon source component, the salt concentration, the nitrogen source component and the concentration, and the composition and the concentration of the phosphorus component, the Listeria monocytogenes was selectively enriched with high efficiency, but the composition which inhibited the growth of Listeria lnokuli was experimented.

Listeria enrichment broth (Merck, Darmstadt, Germany, Listeria Enrichment Broth (LEB) acc. To FDA / IDF-FIL) of the composition shown in Table 1 was used as a control.

The carbon source component selection test was carried out as follows.

(+) - Galactose (Wako, Tokyo, Japan), D (+) - Maltose (YAKURI, Osaka, Japan) 0.25% (g / L) of D-fructose (Sigma-Aldrich, St. Louis, Missouri, USA) and sucrose (Sigma-Aldrich, St. Louis, Missouri, USA) The medium was adjusted to be the same as that of LEB at present and sterilized at 121 캜 for 15 minutes to prepare a medium. After incubation at 35 ± 2 ° C. for 30 hours, the cells were inoculated into ALOA agar (Biomeriux, Marcy L'Etoile, France) for 24 hours at 37 ° C. The cells were inoculated into a medium prepared by mixing Listeria monocytogenes and Listeria monocytogenes A typical colonization of each organism was counted. Listeria enrichment broth (Merck, Darmstadt, Germany) was used as a control for the medium to be developed.

As a result, compared with the case where lactose (Lac), galactose (Gal), xylose, Xyl, maltose, Mal and fructose were used as a carbon source, When sucrose (Suc) was used as a carbon source, Listeria monocytogenes was most abundant, and Merck, Darmstadt, Germany, containing D (+) glucose as a carbon source , Germany Listeria Enrichment Broth (LEB) acc. To FDA / IDF-FIL). On the other hand, it was confirmed that the increase amount of Listeria monocytogenes was not significantly different depending on the carbon source, and that the amount of Listeria monocytogenes was smaller in all carbon sources than in the case of using Listeria monocytogenes from Merck (see )).

In addition, sucrose was selected as a carbon source, and the medium was prepared so as to have concentrations of 0.2%, 0.25% and 0.5% (g / L), and the same test as above was carried out. As a result, at all concentrations, Listeria enrichment broth (LEB) acc. To FDA / IDF-FIL from Merck, Darmstadt, Germany, containing D (+) glucose as a carbon source, )). In particular, it was confirmed that the enrichment amount was large at the concentration of 0.5% (g / L) (FIG. 2 (a-1)).

To determine the nitrogen source component, Proteose Peptone (OXOID, Hampshire, UK) and Beef Extract (Becton Dickinson, CA, USA) were added as nitrogen source components and tested in the same manner as above (FIG. According to the results, Beef Extract was selected as the best nitrogen source, and the concentration was adjusted to 0.6%, 0.8%, and 1.2% (g / L).

As a result, compared with the Listeria enrichment broth (Merck, Darmstadt, Germany, Listeria Enrichment Broth (LEB) acc. To FDA / IDF-FIL) containing yeast extract as a nitrogen source, And 1.2%, respectively. On the other hand, it was confirmed that the increased amount of Listeria monocytogenes was a nitrogen source and that the amount of Listeria monocytogenes was small at all concentrations compared with the case of using Listeria monocytogenes from Merck Co., which contains Yeast Extract (see (c- One)).

In the case of selecting the salt concentration, the medium was prepared with sodium chloride concentrations of 1.5%, 3.0%, 5.0%, 7.0% and 10.0% (g / L)

As a result, Listeria monocytogenes was most abundant at a concentration of 3%, and Listeria enrichment broth (Merck, Darmstadt, Germany, Listeria Enrichment Broth (LEB) acc. To FDA / IDF -FIL), and it was confirmed that even at a concentration of 1.5%, the microbial content was higher than that of Listeria monocytogenes from Merck. On the other hand, it was confirmed that the luciferase activity of Listeria monocytogenes was equal to or less than that in the case of using Listeria broth of Merck, at all concentrations (FIG. 2 (b)).

(Wako, Tokyo, Japan), Di-sodium hydrogen phosphate (Wako, Tokyo, Japan) and Potassium Di-hydrogen phosphate (Wako, Tokyo, Japan) After the preparation, the above test was conducted (Fig. 2 (d)). As a result, potassium di-hydrogen phosphate was selected as the most excellent person.

Then, the medium was prepared by varying the concentrations of potassium dihydrogen phosphate to 0.15%, 0.25%, 0.35%, and 0.5%, and the same test was conducted as above.

As a result, Merck, Darmstadt, Germany, Listeria Enrichment Broth (LEB) acc. To (Merck) containing di-potassium hydrogen phosphate as a ginseng salt at all concentrations above 0.25% FDA / IDF-FIL). On the contrary, it was confirmed that the amount of lysophospholipid of Listeria monocytogenes was small at all concentrations compared with the case of using ginseng salts of Listeria monocytogenes from Merck Co. containing di-potassium hydrogen phosphate (Fig. 2 (d-1)).

Example  3. Listeria Monosaitogenes  Selection Enrichment  Badge Enrichment  effect

Listeria monocytogenes selective enrichment medium was prepared with the ingredients and contents shown in Table 2 below.

The Listeria monocytogenes selective enrichment medium of Table 2 and the Listeria enrichment broth (Merck, Darmstadt, Germany, Listeria Enrichment Broth (LEB) acc. To FDA / IDF-FIL) , Escherichia coli (ATCC 25922), Salmonella enteritidis (ATCC 13076), Shigella sonnei (ATCC 9290), Vibrio parahaemolyticus (ATCC 17803), Yersinia enterocolitica (ATCC 9610), Escherichia coli O157 Bacillus cereus (ATCC 11778), Staphylococcus aureus (ATCC 12600), L. monocytogenes (ATCC 15313), L. innocua (ATCC 43894), Enterobacter aerogenes (ATCC 13048), Citrobacter freundii The cultures were incubated at 35 ± 2 ° C. for 30 hrs and then cultured in ALOA agar (Biomeriux, Marcy L'Etoile, France) at 37 ° C. for 24 hours to obtain a typical colony of each strain Respectively.

In addition, Listeria monocytogenes selective enrichment medium of Table 2 and Listeria monocytogenes broth (Merck, Darmstadt, Germany, Listeria Enrichment Broth (LEB) acc. To FDA / IDF-FIL) And Listeria monocytogenes were mixed at a constant concentration, and incubated at 35 ± 2 ° C. for 30 hours. The cells were cultured in an ALOA agar (Biomeriux, Marcy L'Etoile, France) at 37 ° C. for 24 hours, A typical colony of bacteria was counted.

As a result, as shown in Table 3, when two kinds of Listeria monocytogenes and Listeria monocytogenes were inoculated in the development medium and 12 kinds of strains were inoculated including the two kinds of Listeria monocytogenes and Listeria monocytogenes, 114 To 120%, but the growth of Listeria monocytogenes was rather suppressed. On the contrary, when cultured in Listeria broth (Merck, Darmstadt, Germany, Listeria Enrichment Broth (LEB) acc. To FDA / IDF-FIL) of Merck, no Listeria monocytogenes or Listeria inocua .

Claims (10)

In a Listeria monocytogenes enrichment medium containing a carbon source, a nitrogen source and a phosphorus source,
Comprising sucrose as said carbon source;
Wherein the nitrogen source comprises a beef extract; And
Wherein the phosphorus contains potassium dihydrogen phosphate,
Lt; RTI ID = 0.0 &gt; Listeria monocytogenes &lt; / RTI &gt;
Listeria monocytogenes selective enrichment medium. The Listeria monocytogenes selective enrichment medium according to claim 1, wherein the concentration of sucrose is 0.2 - 0.5%. 2. The Listeria monocytogenes selective enrichment medium according to claim 1, wherein the concentration of the beef extract is 0.6 - 1.8%. The Listeria monocytogenes selective enrichment medium according to claim 1, wherein the concentration of potassium dihydrogen phosphate is 0.25-0.5%. The Listeria monocytogenes selective enrichment medium according to claim 1, wherein the Listeria monocytogenes selective enrichment medium comprises sodium chloride and the concentration of sodium chloride is 0.5-3%. The Listeria monocytogenes selective enrichment medium according to claim 1, wherein the Listeria monocytogenes selective enrichment medium inhibits the growth of Listeria monocytogenes. 2. The Listeria monocytogenes selective enrichment medium according to claim 1, characterized in that the Listeria monocytogenes selective enrichment medium does not affect or inhibits the growth of one or more of the following bacteria:
Escherichia E. coli , Salmonella enteritidis , Shigella sonnei), Vibrio para H. Molina di kusu (Vibrio parahaemolyticus), yeosi California Enterococcus coli urticae (Yersinia enterocolitica), E. coli 0157 (Escherichia E. coli O157), Enterobacter aerogenes , and Citrobacter gram-negative bacteria including freundii ; Or Gram-positive bacteria comprising Bacillus cereus , Staphylococcus aureus , and L. innocua . The method of claim 1, wherein the Listeria monocytogenes selective enrichment medium is selected from the group consisting of tryptone peptone, peptone, sucrose, potassium dihydrogen phosphate, beef extract, acriflavine hydrochloride, cyclohexamide, Lt; RTI ID = 0.0 &gt; l / &lt; / RTI &gt; monocytogenes selective enrichment medium. Culturing a sample in a Listeria monocytogenes selective enrichment medium according to any one of claims 1 to 8 to pre-infect Listeria monocytogenes; And
And detecting Listeria monocytogenes.
Listeria monocytogenes detection method. A Listeria monocytogenesis detection kit comprising the Listeria monocytogenes selective enrichment medium according to any one of claims 1 to 8.

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