JP5968992B2 - Gram-positive cocci detection medium - Google Patents

Description

  The present invention relates to a medium for detecting Gram-positive cocci and a method for detecting Gram-positive cocci using this medium.

  Gram-positive cocci include Staphylococcus bacteria, Streptococcus bacteria, Enterococcus bacteria, and many other species that are regarded as important in a wide range of fields including clinical practice. included.

  Staphylococcus bacteria were previously classified in the Micrococcaceae family, but are now Gram-positive cocci classified in the Bacillales family Staphylococcaceae. Species have been confirmed (Non-Patent Documents 1 and 2). In addition to being resident in human and animal skin, nasal cavity, gastrointestinal tract and other mucosal surfaces, it is widely distributed in nature and mostly non-pathogenic, but some of them are Staphylococcus aureus (Staphylococcus aureus, hereinafter referred to as Staphylococcus aureus), Staphylococcus epidermidis (hereinafter referred to as Staphylococcus epidermidis), Staphylococcus saprophyticus (hereinafter referred to as staphylococcus aureus). Some are pathogenic to humans, such as Methicillin-Resistant Staphylococcus aureus (MRSA), a Staphylococcus aureus that has acquired multidrug resistance, is a major cause of nosocomial infections such as cancer, immunodeficiency, or Serious symptoms may occur when patients with reduced resistance after surgery are infected. Moreover, Staphylococcus epidermidis causes endocarditis and bacteremia due to infection from an indwelling intravascular catheter. Although not toxic compared to MRSA, methicillin-resistant Staphylococcus epidermidis (MRSE), which has acquired multidrug resistance, has also been confirmed. Therefore, it is important to detect these bacteria more quickly and accurately.

  In addition to the clinical field, Staphylococcus aureus is frequently tested in the food field as a major food poisoning bacterium, and is also regarded as important in the veterinary field as a causative bacterium for cattle and chicken diseases. For this reason, detection of Staphylococcus bacteria is widely performed in the clinical, food, environment and animal fields, and a method capable of determining with high sensitivity in a shorter time is desired.

  Among the bacteria belonging to the genus Streptococcus, Streptococcus pyogenes (hereinafter referred to as Streptococcus pyogenes), Streptococcus agalactiae, Streptococcus pneumoniae (hereinafter referred to as Streptococcus pneumoniae, Streptococcus pneumoniae) Pathogens such as stance (Streptococcus mutans), Streptococcus sobrinus (Streptococcus sobrinus) are included (Non-patent Document 2). In particular, Streptococcus pyogenes has the strongest pathogenicity among Streptococcus bacteria, and is a causative bacterium of many diseases such as fulminant hemolytic infection, pharyngitis, tonsillitis, scarlet fever, and glomerulonephritis. Streptococcus pneumoniae, Streptococcus agaractiee are bacterial meningitis and sepsis-causing fungi, but drug-resistant bacteria such as Penicillin-resistant Streptococcus pneumoniae (hereinafter referred to as PRSP) are Staphylococcus. It is regarded as a problem as a causative bacterium of nosocomial infection as well as resistant bacteria of the genus bacteria. Streptococcus mutans and Streptococcus sobrinas are known as the most important causative bacteria of caries, and Streptococcus agalactier is also a causative bacterium of livestock mastitis. Therefore, detection of Streptococcus bacteria is performed in a wide range of fields, and a method capable of determining with high sensitivity in a shorter time is desired.

  The genus Enterococcus was once included in the genus Streptococcus, but is now classified independently as an genus Enterococcus (Non-patent Documents 2 and 3). Permanent in the intestinal tract of humans and animals, and does not exhibit pathogenicity in healthy people, but when infected with patients with reduced immunity, urinary tract infections, postoperative infections of the abdomen, sepsis, meningitis Because it causes endocarditis, it becomes a causative bacterium of nosocomial infections. Enterococcus faecalis, Enterococcus faecium, Enterococcus gallinarum, Enterococcus avium, etc. are clinically isolated Enterococcus faecalis, Enterococcus faecium, Enterococcus gallinarum, and Enterococcus avium. Vancomycin-resistant enterococci (hereinafter referred to as VRE), a drug-resistant bacterium, is often multidrug-resistant and can cause the absence of effective antibiotics. It is regarded as a problem worldwide as a causative fungus. Therefore, in the clinical, environmental, and food fields, a method capable of determining Enterococcus bacteria in a shorter time and with higher sensitivity is desired.

  Usually, these bacteria are detected by detecting Mannit salt medium, Egg York salt medium, Staphylococcus no. It is carried out by a culture method using a selective separation medium based on 110 medium, sheep blood agar medium, brain heart infusion medium and the like. Selective separation medium is a general term for mediums designed so that only microorganisms to be separated can grow as much as possible. It acts to suppress microorganisms that are thought to coexist in a sample, while microorganisms to be separated An antibacterial compound that does not affect the growth of the plant is added as a selective agent. For example, in the case of MRSA, antibiotics such as oxacillin, ceftizoxime, and erythromycin are added to the medium to suppress the growth of methicillin-sensitive bacteria, and polymyxin B to suppress the growth of Pseudomonas aeruginosa, Bacillus subtilis, fungi, and the like. Synoxacin, amphotericin B and the like are added to enhance the selective separation of MRSA (Patent Documents 1, 2, and 3). In addition, in the case of Streptococcus agaractie, also called group B streptococcus, antibiotics such as colistin, nalidixic acid, and gentamicin are added to the culture medium to suppress the growth of Gram-negative rods (non-separating properties). Patent Document 4).

  On the other hand, thiabendazole (2- (4-thiazolyl) benzimidazole), an organic antibacterial substance of benzimidazole type, which is usually used as an antifungal agent, an anthelmintic agent, an agrochemical, etc., is added to the medium to grow fungi And a method for selectively culturing target bacteria (Patent Documents 4 and 5). However, the target bacteria caused by benzimidazole antibacterial substances and imazalyl (1- [2- (allyloxy) -2- (2,4-dichlorophenyl) ethyl] -1H-imidazole), 5,7-dichloro-8-quinolinol, etc. There is no report on the growth promoting effect.

Japanese Patent No. 3298932 JP 2002-034553 A Japanese Patent Publication No. 8-013266 Japanese Patent Laid-Open No. 56-030000 JP 2006-280219 A

Bergey's manual of systematic bacterial second edition, 1, 2001 Yoshiaki Kawamura, Modern Media, 51 (12), 313-327, 2005 Koichi Tanimoto et al., Modern Media, 53 (6), 140-147, 2007 Rie Shibuya et al., Journal of Infectious Diseases, 83 (1), 52-55, 2009

  An object of the present invention is to detect a gram-positive cocci that can promote the growth of a bacterial species to be separated while suppressing the growth of fungi considered to coexist in a sample, and a gram-positive using this medium. The object is to provide a method for detecting cocci.

  As a result of repeated investigations using various selective agents to solve the above problems, the present inventor is considered to coexist in a sample by adding a benzimidazole antibacterial substance and a β-lactam antibiotic to the medium. It was newly found that the growth of Gram-positive cocci to be separated can be promoted while suppressing the growth of fungi. Further, the present inventor has the effect of promoting the growth of Gram-positive cocci to be separated while suppressing the growth of fungi, and imazaril and / or 5,7-dichloro-8-quinolinol and β-lactam antibiotics. The present invention was completed by finding that it was also observed in the added medium.

That is, the present invention has the following configuration.
(1) (a) benzimidazole antibacterial substance and
(B) A medium for detecting Gram-positive cocci characterized by containing a β-lactam antibiotic.
(2) (a) imazalyl and / or 5,7-dichloro-8-quinolinol and
(B) A medium for detecting Gram-positive cocci characterized by containing a β-lactam antibiotic.
(3) The culture medium for detecting Gram-positive cocci according to (1) above, wherein the benzimidazole antibacterial substance is selected from thiabendazole, albendazole and fenbendazole.
(4) The medium for detecting Gram-positive cocci as described in (1) or (3) above, wherein the content of the benzimidazole antibacterial substance is 5 to 5,000 mg / L.
(5) The culture medium for detecting Gram-positive cocci according to any one of (1) to (4) above, wherein the content of β-lactam antibiotic is 1 to 100 mg / L.
(6) The gram positive according to any one of (1) to (5) above, wherein the β-lactam antibiotic is at least one selected from penicillin antibiotics and cephem antibiotics Medium for detection of cocci.
(7) A method of culturing Gram-positive cocci using the medium according to any one of (1) to (6) above.
(8) A method for detecting Gram-positive cocci after culturing by the method according to (7) above.

  According to the present invention, when a benzimidazole antibacterial substance, imazalil and / or 5,7-dichloro-8-quinolinol and β-lactam antibiotics exhibiting antifungal activity are contained in a medium, The growth of Gram-positive cocci can be promoted while inhibiting the growth of possible fungi. Therefore, it is possible to shorten the time required from the start of culture to the result determination as compared with the conventional method. The medium for detecting Gram-positive cocci of the present invention includes MRSA, MSSA (Methicillin-Sensitive Staphylococcus aureus), MRSE, MSSE (Methicillin-Sensitive Staphylococcus epidermidis), PRSP, PSSP (Penicillin-Sensitive Streptococcus pneumoniae), VRE, VSE, -The selective separation medium for each bacterial species, to which a selective agent suitable for each bacterial species to be separated (such as Sensitive Enterococci) is further added, promotes the growth of only the bacterial species to be separated, and the number of bacteria in the sample is small. Even in the case where only it exists, it becomes possible to detect.

  The medium for detecting Gram-positive cocci of the present invention, which can promote the growth of Gram-positive cocci to be separated while suppressing the growth of fungi, is an antifungal substance benzimidazole antibacterial substance, imazalil and / or 5 7-Dichloro-8-quinolinol and β-lactam antibiotics are contained in the medium. In actual culture, an antifungal substance (benzimidazole antibacterial substance, imazalyl and / or 5,7-dichloro-8-quinolinol) and β-lactam antibiotic can be added to the basal medium. In addition, by comparing the growth with a medium containing only β-lactam antibiotics in the basal medium, antifungal substances (benzimidazole antibacterial substances, imazalil and / or 5,7-dichloro-8-quinolinol) and β As a combined effect of lactam antibiotics, a growth promoting action can be confirmed.

  Antifungal substances added in the present invention include thiabendazole, albendazole (methyl 5- (propylthio) -2-benzimidazole carbamate), fenbendazole (methyl 5- (phenylthio) -2-benzimidazole carbamate), carbenda Benzimidazole antibacterial substances such as Jim (methyl 2-benzimidazolecarbamate) and Benomyl (methyl 1- (butylcarbamoyl) -2-benzimidazolecarbamate), Imazaril, 5,7-dichloro-8-quinolinol Although it will not specifically limit if it is at least 1 type containing, The thiabendazole which shows a growth promotion effect | action in a wide concentration range is especially preferable.

  The concentration of the benzimidazole antibacterial substance added in the present invention is a concentration range that promotes the growth of Gram-positive cocci while suppressing the fungal growth by coexisting with the β-lactam antibiotic. As shown in the Examples, since the growth promoting action against gram-positive cocci was observed in the range of 5 to 5,000 mg / L, the benzimidazole antibacterial substance content was in the range of 5 to 5,000 mg / L. It is preferable that

  The β-lactam antibiotic added in the present invention is not particularly limited as long as it is a penicillin or cephem antibiotic. The cephem antibiotic is a general term for antibiotics composed of cephalosporin, cephamycin and oxacephem. Table 1 below shows specific examples of penicillin antibiotics, and Table 2 shows cephem antibiotics. These drugs are usually distributed as salts. In the examples shown later, the abbreviations in parentheses in Tables 1 and 2 were used.

  The β-lactam antibiotic concentration added in the present invention is a concentration range that promotes the growth of gram-positive cocci while suppressing the fungal growth by coexisting with the antifungal substance. As shown in the examples, since the growth promoting action against gram-positive cocci is recognized in the range of 1 to 100 mg / L, the content of β-lactam antibiotics should be in the range of 1 to 100 mg / L. Is preferred.

  The bacterial species to be detected in the present invention is not particularly limited as long as it is a gram-positive cocci. Specific examples are Staphylococcus aureus, Staphylococcus epidermidis, rot staphylococci, Staphylococcus caprae, Staphylococcus haemolyticus, Staphylococcus capitis, Staphylococcus capitis, Staphylococcus capitis Staphylococcus sp ), Streptococcus salivarius, Streptococcus suis and other Streptococcus bacteria, Enterococcus Ekarisu, Enterococcus faecium, Enterococcus Garinaramu, Enterococcus avium, Enterococcus Kaserifurabasu (Enterococcus casseliflavus), Enterococcus Rafinosasu (Enterococcus raffinosus), include Enterococcus bacteria, such as Enterococcus Durance (Enterococcus durans).

  The basal medium used in the present invention includes normal agar medium, blood agar medium, mannitol salt medium, egg yolk salt medium, Staphylococcus no. The medium is not particularly limited as long as it is a medium capable of growing Gram-positive cocci such as 110 medium, sheep blood agar medium, and brain heart infusion medium.

  The gram-positive cocci detection medium of the present invention further contains, as necessary, a chromogenic enzyme substrate suitable for the species to be separated, a selective agent for suppressing growth other than the bacteria to be separated, and other components. It is also possible to add. Usually, the action of the present invention is not affected as long as it is added in the types and concentrations of components that can be used in the selective separation medium.

  The sample used in the present invention is not particularly limited as long as the presence of gram-positive cocci such as those derived from humans or animals, foods, and the environment is suspected.

The form of the detection medium of the present invention is not particularly limited. Any form such as liquid, semi-fluid, solid, and sheet can be used. From the viewpoint of easy detection, a solid medium is preferable, and a flat solid medium is more preferable.
Examples of the solidifying agent for the solid medium include commonly used ones such as agar and agarose.

Examples of the present invention are shown below, but the present invention is not limited to these examples.
The list of test strains used in each example is shown in Table 3 below.

Example 1. Screening of antifungal agents The effects of antifungal agents on the growth of fungi and MRSA in the presence of CFX, a β-lactam antibiotic, were investigated.

(1) Medium preparation The following basic medium components were weighed and suspended in purified water, pH was adjusted, and autoclaved at 121 ° C for 15 minutes.

(2) Addition of β-lactam antibiotic and antifungal action screening substance After autoclaving and cooling to 50 ° C., CFX and antifungal action screening substance were added to the following concentrations. Thereafter, 20 mL of the medium was dispensed into a petri dish and solidified. As a control, a medium supplemented only with CFX was prepared in the same manner.

(3) Bacterial inoculation and culture The pre-cultured strain was suspended in sterile physiological saline to prepare a McFarland No. 1 bacterial solution. Thereafter, a 10-fold serial dilution was prepared, and a bacterial solution diluted 10 ⁰ to 10 ⁻⁶ was added dropwise.
After the dropped bacterial solution was absorbed into the medium, aerobic culture was performed at 35 ° C., and determination was made after 45 hours.

(4) Results Tables 6 to 10 show the results of the action on fungi and MRSA for each additive concentration of each substance. Those in which the growth of the fungus was recognized are indicated by +, and those in which the fungus was not recognized are indicated by-.

  As shown in the above table, antifungal activity was observed in all antifungal activity screening substances. On the other hand, regarding the effect on MRSA, growth enhancement was observed with imazalyl, 5,7-dichloro-8-quinolinol, and thiabendazole.

Example 2 Examination of Thiabendazole Concentration Range The concentration of thiabendazole, which exhibits a growth promoting action on MRSA, was examined by fixing the concentration of CFX, a β-lactam antibiotic.
In all of the following examples up to Example 12, the operations of medium preparation, drug preparation, bacterial solution preparation, and determination were performed in the same manner as in Example 1. In addition, although the data regarding an antifungal action are not shown in the following examples, in all examinations, it was confirmed that Candida tropicalis has an antifungal action.

(1) Basal medium component 17 mg / L of the selective agent was added to the same basic medium component as in Example 1.
(2) Addition of thiabendazole and CFX Thiabendazole and CFX were added so as to have concentrations shown in Table 11 below.

(3) Results The determination results after 24 and 45 hours are shown in Table 12 below.

  As shown in Table 12, growth promoting action was observed for MRSA in the concentration range of thiabendazole 5 to 5,000 mg / L.

Example 3 Examination of β-lactam antibiotic concentration range Additive concentration range that shows growth-promoting action against Staphylococcus aureus and Staphylococcus epidermidis in the presence of thiabendazole using cephamycin CFX or oxacephem LMOX as β-lactam antibiotic I investigated.

(1) Basal medium component The basal medium component similar to Example 1 was used.
(2) Addition of thiabendazole, CFX and LMOX Thiabendazole, CFX and LMOX were added so as to have the concentrations shown in Table 13 below.

(3) Results The determination results after 24 and 45 hours are shown in Tables 14 and 15 below. The results are shown by comparison with a medium without addition of thiabendazole prepared as a control.

  As shown in the above table, the combined effect with thiabendazole shown by comparison with the medium without addition of thiabendazole was observed at the lower limit of 1 mg / L and the upper limit of 100 mg / L for both CFX and LMOX.

Example 4 Examination of combined effects of MPIPC or CZX and thiabendazole Using penicillin MPIPC or cephalosporin CZX as β-lactam antibiotics, we investigated the growth-promoting action against MRSA and Staphylococcus epidermidis in the presence of thiabendazole.

(1) Basal medium components Using the following basic medium components, autoclaved at 121 ° C for 15 minutes, cooled to 50 ° C, and then added sheep blood.

(2) Addition of thiabendazole, MPIPC, and CZX Thiabendazole, MPIPC, and CZX were added so as to have concentrations shown in Table 17 below.

(3) Results The determination results after 24 and 45 hours are shown in Tables 18 and 19 below. The results are shown by comparison with a medium without addition of thiabendazole prepared as a control.

  As shown in the above table, the combined effect with thiabendazole shown by comparison with the medium without thiabendazole addition was observed for both MPIPC and CZX.

Example 5 FIG. Examination of the action of Staphylococcus spp.
The presence or absence of a growth promoting effect on MRSA, Staphylococcus epidermidis, Staphylococcus caprae, Staphylococcus hemolyticus, Staphylococcus capitis, Staphylococcus squirrel was examined.

(1) Basal medium component The basal medium component similar to Example 1 was used.
(2) Addition of thiabendazole and CFX Thiabendazole and CFX were added so as to have concentrations shown in Table 20 below.

(3) Results The determination results after 24 and 45 hours are shown in Tables 21 to 26 below. The results are shown by comparison with a medium without addition of thiabendazole prepared as a control.

  As shown in the above table, the growth-promoting effect of the combined use of thiabendazole and β-lactam antibiotics was observed in all the species examined.

Example 6 Examination using normal agar medium The presence or absence of growth-promoting action on MRSA was examined when ordinary agar medium was used as the basic medium.

(1) Preparation of basal medium A basal medium was prepared according to the composition table of the normal agar medium shown below.

(2) Addition of thiabendazole and CFX Thiabendazole was added at 80 mg / L and CFX at 5 mg / L.

(3) Results The determination results after 24 and 45 hours are shown in Table 28 below. The results are shown by comparison with a medium without addition of thiabendazole prepared as a control.

  As shown in Table 28, a growth promoting effect on MRSA was also observed when a normal agar medium was used as the basal medium.

Example 7 Examination of shortening of culture time The growth promotion effect | action with respect to MRSA by adding thiabendazole and CFX was investigated by setting culture time as 16, 18, and 20 hours.

(1) Basal medium component The basal medium component similar to Example 1 was used.
(2) Addition of thiabendazole and CFX Thiabendazole and CFX were added to the concentrations shown in Table 29 below.

(3) Results The determination results after 16, 18, and 20 hours are shown in Table 30 below.

  As shown in Table 30, a growth promoting action was observed up to 2,000 mg / L of thiabendazole even in the determination after 16 hours of culture. From these results, it was suggested that the medium supplemented with the agent of the present invention can detect a small number of bacteria in a shorter time.

Example 8 FIG. Examination of the combined effect of CFX and albendazole Using albendazole as a benzimidazole antibacterial substance, we investigated the growth promoting action against MRSA in the presence of CFX.

(1) Basal medium component The basal medium component similar to Example 1 was used.
(2) Addition of albendazole and CFX Albendazole (ABZ) was added at 80 mg / L and CFX at 5 mg / L.

(3) Results The determination results after 24 and 45 hours are shown in Table 31 below. The results are shown by comparison with a medium without addition of albendazole prepared as a control.

  As shown in Table 31, even when albendazole was used as the benzimidazole antibacterial substance, a growth promoting action against MRSA was observed in combination with CFX.

Example 9 Examination of the combined effect of CFX and fenbendazole Using fenbendazole as a benzimidazole antibacterial substance, we investigated the growth promoting action against MRSA in the presence of CFX.

(1) Basal medium component The basal medium component similar to Example 1 was used.
(2) Addition of fenbendazole and CFX Fenbendazole (FBZ) was added at 80 mg / L and CFX at 5 mg / L.

(3) Results The determination results after 24 and 45 hours are shown in Table 32 below. The results are shown by comparison with a medium without addition of fenbendazole prepared as a control.

  As shown in Table 32, even when fenbendazole was used as a benzimidazole antibacterial substance, a growth promoting action against MRSA was observed in combination with CFX.

Example 10 Examination of the effect of Streptococcus bacteria on each bacterial species The presence or absence of a growth promoting action against Streptococcus pyogenes, Streptococcus agalactier, and Streptococcus pneumoniae by the combined use of thiabendazole and β-lactam antibiotics was examined.

(1) Basal medium component The basal medium component similar to Example 4 was used.
(2) Addition of thiabendazole, CFX, and LMOX Thiabendazole, CFX, and LMOX were added so as to have concentrations shown in Table 33 below.

(3) Results The determination results after 24 and 45 hours are shown in Tables 34 to 36 below. The results are shown by comparison with a medium without addition of thiabendazole prepared as a control.

  As shown in the above table, the growth-promoting effect of the combined use of thiabendazole and β-lactam antibiotics was observed in all the species examined.

Example 11 Examination of the combined effect of CFX and albendazole Using albendazole as a benzimidazole antibacterial substance, we investigated the growth-promoting effect on Streptococcus agaractiee in the presence of CFX.

(1) Basal medium component The basal medium component similar to Example 4 was used.
(2) Addition of albendazole and CFX Albendazole was added at 80 mg / L and CFX at 3 mg / L.

(3) Results The determination results after 24 and 45 hours are shown in Table 37 below. The results are shown by comparison with a medium without addition of albendazole prepared as a control.

  As shown in Table 37, even when albendazole was used as a benzimidazole antibacterial substance, a growth promoting action against Streptococcus agalactier was observed in combination with CFX.

Example 12 Examination of the combined effect of CFX and fenbendazole Using fenbendazole as a benzimidazole antibacterial substance, the growth promoting effect on Streptococcus agaractie was investigated in the presence of CFX.

(1) Basal medium component The basal medium component similar to Example 4 was used.
(2) Addition of fenbendazole and CFX Fenbendazole was added at 80 mg / L and CFX at 3 mg / L.

(3) Results The results of determination after 24 and 45 hours are shown in Table 38 below. The results are shown by comparison with a medium without addition of fenbendazole prepared as a control.

  As shown in Table 38, even when fenbendazole was used as a benzimidazole antibacterial substance, a growth promoting action against Streptococcus agalactier was observed in combination with CFX.

  The medium for detecting Gram-positive cocci according to the present invention contains a benzimidazole antibacterial substance that exhibits antifungal activity, imazalil and / or 5,7-dichloro-8-quinolinol and a β-lactam antibiotic in the medium. While suppressing the growth of fungi considered to coexist in the sample, the growth of Gram-positive cocci can be promoted, and the time required from the start of culture to the result determination can be shortened compared to the conventional method. Therefore, it is useful for detection of various gram-positive cocci in a wide range of fields including clinical, food, environment, and animals.

Claims (14)

(A) benzimidazole antibacterial substance 5-5,000 mg / L and
(B) 1-100 mg / L of β-lactam antibiotic
Containing,
The benzimidazole antibacterial substance is selected from thiabendazole, albendazole and fenbendazole;
The β-lactam antibiotic is at least one selected from penicillin antibiotics, cephalosporin antibiotics, cephamycin antibiotics and oxacephem antibiotics;
While suppressing the growth of Candida tropicalis (Candida tropicalis), to promote the growth of bacterial strains to be separated, and,
The species to be separated are Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus caprae, Staphylococcus haemolyticus, Staphylococcus capitotis (Staphylococcus haemolyticus) Selected from Staphylococcus capitis, Staphylococcus sciuri, Streptococcus pyogenes, Streptococcus agalactiae and Streptococcus pneumoniae ,
A culture medium for detecting bacterial species to be separated , characterized by The benzimidazole antibacterial is thiabendazole , the penicillin antibiotic is oxacillin, the cephalosporin antibiotic is ceftizoxime, the cephamycin antibiotic is cefoxitin, and the oxacephem antibiotic is detection medium according to claim 1, characterized in that the latamoxef. The benzimidazole antibacterial substance is albendazole and / or fenbendazole, the β-lactam antibiotic is cefoxitin, and the species to be isolated is S. aureus or Streptococcus agalactier The detection medium according to claim 1. The medium for detection according to claim 3, wherein the concentration of the benzimidazole antibacterial substance is 80 to 5,000 mg / L. (A) Imazaryl 10-50 mg / L and / or 5,7-dichloro-8-quinolinol 12-15 mg / L and
(B) Cefoxitin 1-100 mg / L
Containing,
Promoting the growth of the species to be isolated while suppressing the growth of Candida tropicalis , and
The bacterial species to be separated is Staphylococcus aureus ,
A culture medium for detecting bacterial species to be separated , characterized by (A) benzimidazole antibacterial substance 5-5,000 mg / L and
  (B) 1-100 mg / L of β-lactam antibiotic
  Using a medium containing
  The benzimidazole antibacterial substance is selected from thiabendazole, albendazole and fenbendazole;
  The β-lactam antibiotic is at least one selected from penicillin antibiotics, cephalosporin antibiotics, cephamycin antibiotics and oxacephem antibiotics;
  Promoting the growth of the species to be isolated while suppressing the growth of Candida tropicalis, and
  The method for promoting the growth of a bacterial species to be separated, characterized in that the bacterial species to be separated is a bacterium belonging to the genus Staphylococcus or Streptococcus. That the benzimidazole antibacterial substance is thiabendazole,
  The penicillin antibiotic is oxacillin, the cephalosporin antibiotic is ceftizoxime, the cephamycin antibiotic is cefoxitin, and the oxacephem antibiotic is latamoxef,
  The Staphylococcus bacterium is selected from Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus caprae, Staphylococcus hemolyticus, Staphylococcus capitis and Staphylococcus schiuli, and
  The Streptococcus bacterium is selected from Streptococcus pyogenes, Streptococcus agalactier and Streptococcus pneumoniae,
  The growth promoting method according to claim 6, wherein: The benzimidazole antibacterial substance is albendazole and / or fenbendazole, the β-lactam antibiotic is cefoxitin, and the species to be isolated is S. aureus or Streptococcus agalactier The growth promoting method according to claim 6. The method according to claim 8, wherein the concentration of the benzimidazole antibacterial substance is 80 to 5,000 mg / L. (A) Imazaryl 10-50 mg / L and / or 5,7-dichloro-8-quinolinol 12-15 mg / L and
  (B) Cefoxitin 1-100 mg / L
  Using a medium containing
  Promoting the growth of the species to be isolated while suppressing the growth of Candida tropicalis, and
  The bacterial species to be separated is a Staphylococcus bacterium,
  A method for promoting the growth of bacterial species to be separated, characterized by A medium for use in the growth promoting method according to any one of claims 6 to 10. A method for detecting a bacterial species to be separated, after promoting the growth of the bacterial species to be separated by the growth promoting method according to any one of claims 6 to 10 . A method for culturing the bacterial species to be separated using the detection medium according to any one of claims 1 to 5 or the medium according to claim 11 . A method for detecting a bacterial species to be separated after culturing the bacterial species to be separated using the detection medium according to any one of claims 1 to 5 or the medium according to claim 11. .