JP2003310298A - Coloring medium for discriminating candida - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a coloring medium for discriminating Candida, having excellent discrimination ability in discrimination i.e., whether Candida albicans, Candida tropicalis or both are present, in a short time by color of a colony on the medium, wherein, by using the medium, the Candida albicans or the Candida tropicalis from another fungus belonging to the Candida is clearly discriminated by color tone on the medium. <P>SOLUTION: The coloring medium for discriminating Candida albicans contains magenta-gel as a chromogenic enzyme substrate and the coloring medium for discriminating Candida tropicalis contains x-glc. <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a coloring medium for distinguishing Candida that can easily distinguish Candida albicans, which is a causative bacterium of candidiasis, from Candida tropicalis.

[0002]

2. Description of the Related Art Infection status, treatment status in candidiasis,
In diagnosing the recurrence situation, it is clinically important to confirm the number of bacteria along with the morphology, tissue lesions, and gross inflammatory findings of Candida by microscopic examination. In order to confirm the number of such bacteria, a culture method is widely used, and various media such as Mizuno-Takada medium and Sabouraud medium have already been marketed. All of these media contain a carbon source, a nitrogen source, and a substance that suppresses the growth of general bacteria, and the purpose is to detect only Candida.

In using these media, after culturing the specimen collected from the affected area in the media for about 2 to 3 days,
The present condition of candidiasis is diagnosed by observing the growth of bacteria and the number and morphology of colonies with the naked eye. However, in the conventional medium, since it takes 2 or 3 days to culture the bacteria, it is impossible to make a quick diagnosis and there is a problem that a quick treatment cannot be started.

Therefore, an aqueous solution containing at least an amino acid, a peptide, a saccharide, and a bacterial growth inhibiting substance is mixed with a pH indicator which changes its color in an acidic range where the growth of various bacteria is suppressed.
Similarly, a medium for candidiasis diagnosis has been proposed in which candidiasis can be easily diagnosed in a short time without requiring skill by using a medium mixed in a medium whose pH is adjusted so that various bacteria are suppressed ( Japanese Patent Publication No. 46115/1989).

On the other hand, with respect to candidiasis, Candida albicans is known as a major causative agent, but other than this, Candida glabrata is the causative agent and Candida tropicalis is the causative agent. Things are increasing.

[0006] Of these Candida, Candida albicans and Candida glabrata can be easily distinguished by the color tone of the colonies on the medium, and by utilizing this, the pathogenic bacterium of candidiasis can be easily and quickly determined. , A medium for simple identification of Candida that can establish therapeutic measures at an early stage has been proposed (JP-A-6-78793 and JP-A-6-7873).
277094).

[0007]

However, Candida albicans and Candida tropicalis not only have similar morphological characteristics, but also have no known differences in biochemical characteristics. There was a problem that it was indistinguishable at all.

Therefore, the present invention has been made by paying attention to such a conventional problem, and can easily and quickly develop Candida albicans and Candida tropicalis without requiring skilled techniques. An object is to provide a chromogenic medium for distinguishing Candida that can be discriminated.

[0009]

Means for Solving the Problems The present inventor
As a result of intensive research to develop a medium for discriminating between Albicans and Candida tropicalis, magenta-gal and x-glc
By adding C.
The present inventors have completed the present invention by finding that Albicans and Candida tropicalis can be easily distinguished.

That is, the present invention has the following structure. (1) A coloring medium for distinguishing Candida albicans, which contains magenta-gal as a chromogenic enzyme substrate. (2) The coloring medium for distinguishing Candida albicans according to (1), wherein the magenta-gal content is in the range of 0.1 to 5 g / L. (3) The coloring medium for distinguishing Candida albicans according to (1) or (2), which further contains glucose. (4) The coloring medium for distinguishing Candida albicans according to (1) to (3), wherein the glucose content is in the range of 1 to 15 g / L. (5) Containing 1 to 5 g of magenta-gal, 1 to 30 g of peptone, 0.5 to 6.0 g of yeast extract, 1 to 15 g of glucose, and 5 to 20 g of agar per 1 L of the medium (1) to (4) A coloring medium for Candida albicans differentiation. (6) A chromogenic medium for distinguishing Candida tropicalis, which contains x-glc as a chromogenic enzyme substrate. (7) The coloring medium for distinguishing Candida tropicalis according to (6), wherein the content of x-glc is in the range of 0.1 to 5 g / L. (8) The coloring medium for distinguishing Candida tropicalis according to (6) to (7), which further contains saccharose. (9) The coloring medium for Candida tropicalis discrimination according to (6) to (8), wherein the sucrose content is in the range of 1 to 15 g / L. (10) x-glc 0.1 to 5 g, peptone 1 to 30 g, yeast extract 0.5 to 6.0 g, sucrose 1 to 15 g, and agar 5 to 20 g per 1 L of the medium (6) to (9) Candida tropicalis chromogenic medium for differentiation. (11) A coloring medium for distinguishing Candida albicans and / or Candida tropicalis, which contains magenta-gal and x-glc as chromogenic enzyme substrates.

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in more detail below. The characteristic of the coloring medium for Candida albicans discrimination of the present invention is that magenta-ga is used as a coloring enzyme substrate in the medium.
l is included. This chromogenic enzyme substrate is decomposed by galactosidase produced by Candida albicans, and turns purple in the medium. Therefore, when Candida albicans grows on the medium, it is visible (visual).
A purple colony is observed.

The amount of the chromogenic enzyme substrate blended is 0.1 to 5
It is preferably in the range of g / L, and more preferably,
It is in the range of 0.5 to 1 g / L. Substrate content is 0.1
If it is less than g / L, it takes time to discolor the colony, and the detection sensitivity is lowered. On the contrary, there are also Candida albicans that do not grow when the amount exceeds 5 g / L.

In the present invention, the discoloration of colonies becomes more distinct by adding glucose, which is a nutrient source, in the medium in addition to magenta-gal which is the chromogenic enzyme substrate. That is, the discriminating power can be further improved as compared with the case where both the color-developing enzyme substrate and glucose are combined and the enzyme substrate is used alone.

The glucose content is preferably in the range of 1 to 15 g / L, more preferably 2 to 10 g / L. When the glucose content is less than 1 g / L, the growth of Candida albicans becomes extremely poor. Conversely, 1
When it exceeds 5g / L, magenta-gal which is a chromogenic enzyme substrate
Will not be decomposed and the colonies will not develop color.

The coloring medium for distinguishing Candida albicans of the present invention contains at least 1 L of the medium as a nutrient component,
It is preferable to contain 0.1 to 5 g of magenta-gal, 1 to 30 g of peptone, 0.5 to 6.0 g of yeast extract, 1 to 15 g of glucose, and 5 to 20 g of agar. In addition to these nutrients, nitrogen sources such as ammonium sulphate, sugars such as maltose and sucrose, and optionally other minerals and vitamins may be included as nutrient sources that promote the growth of Candida albicans. You can

As an antibacterial agent for suppressing the growth of various bacteria, there are aminoglycoxide type antibiotics such as gentamicin, streptomycin, kanamycin and neomycin, a wide range of antibiotics such as chloramphenicol and tetracycline, and a wide range of antibacterial spectrum. Examples include penicillin and cephalosporin, with chloramphenicol being preferred.

The medium of the present invention can take any form of liquid, semi-fluid and solid, but from the viewpoint of ease of detection and the like,
Solid media are preferred, and more preferably in the form of plate solid media. Examples of the solidifying agent for the solid medium include those commonly used such as agar, carrageenan, and agarose.

The present invention also provides x-gl as a chromogenic enzyme substrate.
Provided is a coloring medium for distinguishing Candida tropicalis, which contains c. This chromogenic enzyme substrate is decomposed by β-glucosidase produced by Candida tropicalis and turns light green in the medium.

The amount of the color-developing enzyme substrate blended is 0.1 to 5
It is preferably in the range of g / L, and more preferably,
It is in the range of 0.1 to 3 g / L. Substrate content is 0.1
If it is less than g / L, it takes time for the color change of the colony. On the contrary, if it exceeds 5 g / L, some Candida tropicalis does not grow.

In the present invention, by adding saccharose as a nutrient source in the medium in addition to x-glc which is the above-mentioned chromogenic enzyme substrate, it is possible to change the color from light green to green, and to further improve the color tone of colonies. Becomes clear and the identification of Candida tropicalis becomes easy.

The amount of sucrose blended is 1 to 15 g / L.
Is preferable, and more preferably 2 to 10 g / L. If the amount of sucrose added is less than 1 g / L, the growth of Candida tropicalis will be extremely poor.
On the contrary, when it exceeds 15 g / L, x-gl which is a chromogenic enzyme substrate
The c is not decomposed and the colonies no longer develop color.

The coloring medium for distinguishing Candida tropicalis of the present invention contains at least 1 L of the medium as a nutrient,
It preferably contains 0.1 to 5 g of x-glc, 1 to 30 g of peptone, 0.5 to 6.0 g of yeast extract, 1 to 15 g of sucrose, and 5 to 20 g of agar. These other optional components can also be included in the same manner as the Candida albicans differentiation coloring medium.

Furthermore, the present invention uses ma as a chromogenic enzyme substrate.
It is possible to provide a coloring medium for distinguishing Candida albicans and / or Candida tropicalis characterized by containing genta-gal and x-glc. Thus, by including a chromogenic enzyme substrate that distinguishes both Candida albicans and Candida tropicalis, even if both are present, their color tones will be different, so it is easy to distinguish them. You can

[0024]

INDUSTRIAL APPLICABILITY The present invention is excellent in distinctiveness when judging by the color tone of a colony on a medium whether Candida albicans is present, Candida tropicalis is present, or both are present. In addition, it is possible to provide a coloring medium for Candida differentiation that can be distinguished in a short time. Therefore, according to the present invention, considering that the isolation frequency of Candida tropicalis is higher than that of Candida albicans, both strains can be differentially identified, which is considered to be clinically significant.

[0025]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto.

Example 1. Examination of Coloring Substances of Candida Colony 4.5 g of the medium components shown in Table 1 were weighed and dissolved in 100 mL of purified water. After dissolution, the pH was adjusted to 6.4 ± 0.2, the respective reagents shown in Table 2 were added in the specified amounts shown in Table 3, and autoclaved at 121 ° C. for 15 minutes. After cooling to 50 ° C., each 20 mL was dispensed into a petri dish and solidified. After solidification, it was dried for 30 minutes. In addition, (12) -shown in Table 3
The reagent of (14) (tetrazolium salt) cannot be autoclaved, so only the medium was autoclaved. After cooling the medium to 50 ° C., a reagent which had been sterilized by filtration in advance was added to the medium.

Table 1 Composition of basal medium ──────────────── Casein peptone 2.5g Yeast extract powder 5.0g Glucose 5.0g Sucrose 5.0 g Trehalose 5.0g Agar 15.0g ──────────────── pH 6.4 ± 0.2

[0028] Table 2 Additional reagents ──────────────────────────── (1) Copper (II) sulfate pentahydrate (2) Copper nitrate trihydrate (3) Zinc sulfate heptahydrate (4) Cobalt nitrate hexahydrate (5) Barium chromate (6) Sodium tungstate dihydrate (7) Thallium acetate (I) (8) Anhydrous aluminum sulfate (9) Potassium tellurite (10) Ammonium molybdate tetrahydrate (11) Iron (III) citrate n hydrate (12) TTC: 2,3,5-Triphenyltetrazolium Chloride (13) TTV: Tetrazolium Violet (14) TTB: Trtrazolium Blue (15) ONPG: o-nitrophenyl-β-D-galactopyranoside (16) ONXY: o-nitrophenyl-β-D-xyopyranoside (17) x-gal (18) x-glu (19) magenta-gal (20) salmon-gal (21) x-glu (22) L-proline (23) nit-galactosaminide ────────────────────────────

[0029] Table 3 Addition amount of each reagent (1000 mL) ─────────────────────── Metal salt (1)-(11) 2.0 g 0.01 g of tetrazolium salt (12)-(14) Chromogenic substrate (15) to (22) 0.1 g ───────────────────────

The following 5 strains (A) to (E) were subcultured in advance on the medium prepared above with a commercially available potato dextrose agar medium, and the obtained strain was smeared and inoculated with a platinum loop (10 μL). And cultured at 37 ° C. for 3 days. The results are shown in Table 4. (A): Candida albicans 3092 (B): Candida glabrata 3388 (C): Candida tropicalis 3395 (D): Candida parapsilosis 3169 (E): Candida krusei 3389

[0031] Table 4 Coloring of Candida colonies when each reagent was added ──────────────────────────────────── ─                         (A) (B) (C) (D) (E) ──────────────────────────────────── ─ 1) Copper sulfate brown colorless brown brown yellow brown 2) Copper nitrate brown colorless brown brown brown 3) Zinc sulfate colorless colorless colorless colorless colorless 4) Cobalt nitrate colorless, not developing brown brown brown 5) Barium chromate colorless colorless colorless colorless colorless colorless 6) Sodium tungstate                           Colorless colorless colorless colorless colorless 7) Thallium acetate No growth No growth No growth No growth No growth Without 8) Aluminum sulfate colorless colorless colorless colorless colorless 9) Potassium tellurite black black black yellow brown gray black 10) Ammonium molybdate                           Yellow colorless brown brown brown 11) Iron citrate Colorless Colorless Colorless Colorless Colorless 12) TTC crimson colorless colorless crimson red colorless 13) TTV Crimson colorless Crimson Crimson pink 14) TTB colorless, colorless, colorless, colorless, colorless 15) ONPG yellow colorless colorless yellow colorless 16) ONXY yellow colorless yellow yellow colorless 17) x-gal colorless colorless colorless colorless colorless 18) x-glc colorless colorless green colorless colorless 19) magenta-gal light purple colorless colorless achromatic colorless colorless 20) salmon-gal colorless colorless colorless colorless colorless 21) x--glu colorless colorless colorless colorless colorless 22) L-proline yellow colorless yellow colorless colorless 23) nit-galactosaminide yellow colorless colorless yellow colorless colorless ──────────────────────────────────── ─

As is clear from the results shown in Table 4, in the metal salt, copper (II) sulfate pentahydrate, cobalt nitrate hexahydrate, and aluminum molybdate tetrahydrate turned brown, and potassium tellurite. There was something that turned black. However, in each case, a plurality of bacteria had the same color, and the strain could not be discriminated. Although the tetrazolium salts of TTC and TTV turned red, the strains could not be discriminated in the same manner as the metal salts, because they showed the same color in a plurality of bacteria. On the other hand, in the chromogenic substrate, magenta-gal tended to be colored only by Candida albicans, and x-glc also showed Candida albicans.
Only the tropicalis tended to be colored.

Example 2 Examination of Sugar Addition A color-forming substrate that reacts with a glycolytic enzyme produced by Candida can suppress color development when a large amount of sugar is contained in the medium components. On the other hand, however, sufficient sugar is required for the growth of Candida, and if the sugar component is too small, the growth will be poor.
If the growth is poor, the production of degrading enzymes also becomes poor, and the chromogenic substrate cannot be decomposed, and no color is seen. Therefore, the optimum amount of growth and color development was examined using sugars used by C. albicans and C. tropicalis. The following six types of sugar are commonly used. (1) Glucose (2) Saccharose (3) Trehalose (4) Galactose (5) Xylose (6) Maltose

In the basal medium used in Example 1 from which sugar components were removed, 0 g of each sugar of the above (1) to (6) and 2.0
The presence or absence of color development was confirmed by adding g, 4.0 g, 6.0 g, 8.0 g, 10.0 g / L and adding a color-developing substrate at 0.2 g / L. The results are shown in Tables 5 and 6. As is clear from Table 5, when magenta-gal is used as the chromogenic enzyme substrate, only Candida albicans develops from purple to light purple in the presence of glucose. On the other hand, as is clear from Table 6, when x-glc is used, only Candida tropicalis develops from light green to green in the presence of saccharose.

Table 5 When magenta-gal was used ────────────────────────────────────── Glucose 0 2 4 6 8 10g / L colony C.albicans3092 diameter 0.5 1.5 2.5 2.5 2.5 2.5 mm color colorless purple purple purple light purple light purple light purple C.tropicalis3395 diameter 0.5 2.0 3.5 3.5 4.0 4.0 color colorless colorless colorless colorless colorless colorless colorless ────── ─────────────────────────────── Sucrose 0 2 4 6 8 10 g / L colony C.albicans 3092 diameter 0.5 0.5 1.0 1.0 1.0 1.0 mm color colorless colorless colorless colorless colorless colorless C. tropicalis3395 diameter 0.5 2.0 3.5 3.5 4.0 4.0 color colorless colorless colorless colorless colorless colorless ────────────────────────── ─────────── Trehalose 0 2 4 6 8 10 g / L Colony C. albicans 3092 Diameter 0.5 1.5 2.5 2.5 2.5 2.5 2.5 mm color colorless colorless colorless colorless colorless colorless C. tropicalis3395 diameter 0.5 2.0 3.0 3.0 4.0 4.0 color colorless colorless colorless colorless colorless colorless ────────────────────────── ─────────── Galactose 0 2 4 6 8 10 g / L colony C.albicans3092 diameter 0.5 1.0 1.0 1.0 1.5 1.5 mm color colorless colorless colorless colorless colorless colorless colorless C.tropicalis 3395 diameter 0.5 2.0 3.5 3.5 4.0 4.0 colored Colorless Colorless Colorless Colorless Colorless Colorless Colorless ──────────────────────────────────── Xylose 0 2 4 6 8 10 g / L colony C.albicans3092 diameter 0.5 1.5 2.0 2.5 2.5 2.5 mm color colorless colorless colorless colorless colorless colorless colorless C.tropicalis3395 diameter 0.5 2.0 2.0 2.0 2.0 2.0 color colorless colorless colorless colorless colorless colorless ────────────── ────────────────────── ─ Maltose 0 4 6 8 10g / L colonies C.albicans3092 diameter 0.5 1.0 1.0 1.0 1.5 1.5 mm Color Colorless Colorless Colorless Colorless Colorless Colorless C.tropicalis3395 diameter 0.5 1.5 2.0
2.0 2.0 2.0 ──────────────────────────────────── ─

[0036] Table 6 When using X-glu ──────────────────────────────────── ─   Glucose 0 2 4 6 8 10 g / L                Colony C. albicans 3092 Diameter 0.5 1.5 2.5 2.5 2.5 2.5 mm                Color Colorless Colorless Colorless Colorless Colorless Colorless C. tropicalis3395 Diameter 0.5 2.0 3.5 3.5 4.0 4.0                 Color Colorless Colorless Colorless Colorless Colorless Colorless   ─────────────────────────────────── ─ Sucrose 0 2 4 6 8 10 g / L                Colony C.albicans 3092 Diameter 0.5 0.5 1.0 1.0 1.0 1.0 mm                Color Colorless Colorless Colorless Colorless Colorless Colorless C. tropicalis3395 Diameter 0.5 2.0 3.5 3.5 4.0 4.0                 Color colorless light green green green green green   ─────────────────────────────────── ─   Trehalose 0 2 4 6 8 10 g / L                Colony C. albicans 3092 Diameter 0.5 1.5 2.5 2.5 2.5 2.5 mm                Color Colorless Colorless Colorless Colorless Colorless Colorless C. tropicalis3395 Diameter 0.5 2.0 3.0 3.0 4.0 4.0                 Color Colorless Colorless Colorless Colorless Colorless Colorless   ─────────────────────────────────── ─  Galactose 0 2 4 6 8 10 g / L                colony C.albicans 3092 Diameter 0.5 1.0 1.0 1.0 1.5 1.5 mm                Color Colorless Colorless Colorless Colorless Colorless Colorless C. tropicalis3395 Diameter 0.5 2.0 3.5 3.5 4.0 4.0                 Color Colorless Colorless Colorless Colorless Colorless Colorless   ─────────────────────────────────── ─  Xylose 0 2 4 6 8 10 g / L                Colony C. albicans 3092 Diameter 0.5 1.5 2.0 2.5 2.5 2.5 mm                Color Colorless Colorless Colorless Colorless Colorless Colorless C. tropicalis3395 diameter 0.5 2.0 2.0 2.0 2.0 2.0                 Color Colorless Colorless Colorless Colorless Colorless Colorless   ─────────────────────────────────── ─   Maltose 0 2 4 6 8 10 g / L                Colony C.albicans 3092 Diameter 0.5 1.0 1.0 1.0 1.5 1.5 mm                Color Colorless Colorless Colorless Colorless Colorless Colorless C. tropicalis3395 Diameter 0.5 1.5 2.0 2.0 2.0 2.0                 Color Colorless Colorless Colorless Colorless Colorless Colorless   ─────────────────────────────────── ─

Example 3 Preparation of Main Medium After weighing the raw materials having the following composition, purified water was added to bring the total amount to 100.
It was made 0 mL and sterilized at 100 ° C. for 20 minutes. After sterilization, 50 ℃
After cooling to 20 mL, pour 20 mL each into a petri dish, solidify, and
This was dried for a minute to obtain the main medium.

Table 7 Composition Main medium (1000 mL) ───────────────────── Casein peptone 15g Yeast extract 3g Glucose 4g Sucrose 4g Trehalose 1g Zinc sulfate 0.2g Aluminum sulfate 0.2g Agar 14g magenta-gal 0.4g x-glu 0.3g IPTG 0.5g Chloramphenicol 0.05g ───────────────────── pH 6.4 ± 0.2 IPTG: Isopropyl-β-D-thiogalactopyranoside

This medium was inoculated with 1 platinum loop (10 μL) of a bacterium that had been subcultured in advance on a potato dextrose agar medium, and cultured at 37 ° C. for up to 3 days. Judgment is the first day,
I went on the second and third days. The results are shown in Tables 8 to 10. As is clear from the table, both Candida albicans and Candida tropicalis develop light purple and green from the first day of culture, respectively, and other fungi do not develop. This means that by using this medium, Candida albicans and Candida tropicalis can be quickly distinguished from each other without requiring a special technique.

[0040]  Table 8 ──────────────────────────────────── ─   Day 1 Colony                                     Diameter (mm) Color ──────────────────────────────────── ─    Candida albicans 3092 1.0 light purple Candida albicans 3093 1.0 light purple Candida albicans 3171 1.0 light purple Candida albicans 3536 1.0 light purple Candida tropicalis 3395 1.5 green Candida tropicalis 3172 1.0 green Candida krusei 3170 1.0 colorless Candida krusei 3389 1.0 colorless Candida glabrata 3388 0.5 colorless Candida glabrata 3390 0.5 colorless Candida parapsilosis 3091 0.5 colorless Candida parapsilosis 3169 0.5 colorless Candida guilliermondii 3426 0.5 0.5 colorless Candida famata 3777 0.5 0.5 colorless Trichosporon cutaneum 3455 0.5 colorless Trichosporon cutaneum 3456 0.5 colorless Trichosporon beigelii 3779 0.5 0.5 colorless Trichosporon beigelii 3780 0.5 colorless Esherichia coli 675 NG- Citrobacter freundii 389 NG- Proteus mirabilis 436 NG- Enterobacter cloacae 606 NG- Pseudomonas aeruginosa 1376 NG- ──────────────────────────────────── ── NG: shows that colony growth was not observed.

[0041]  Table 9 ──────────────────────────────────── ── Day 2 Colony                                     Diameter (mm) Color ──────────────────────────────────── ── Candida albicans 3092 2.0 purple Candida albicans 3093 2.0 purple Candida albicans 3171 2.0 purple Candida albicans 3536 2.0 Purple Candida tropicalis 3395 3.0 green Candida tropicalis 3172 3.0 green Candida krusei 3170 2.5 colorless Candida krusei 3389 2.0 colorless Candida glabrata 3388 1.0 colorless Candida glabrata 3390 1.0 colorless Candida parapsilosis 3091 1.0 colorless Candida parapsilosis 3169 1.0 colorless Candida guilliermondii 3426 1.2 colorless Candida famata 3777 1.5 colorless Trichosporon cutaneum 3455 1.0 colorless Trichosporon cutaneum 3456 1.0 colorless Trichosporon beigelii 3779 0.7 colorless Trichosporon beigelii 3780 0.7 colorless Esherichia coli 675 NG- Citrobacter freundii 389 NG- Proteus mirabilis 436 NG- Enterobacter cloacae 606 NG- Pseudomonas aeruginosa 1376 NG- ──────────────────────────────────── ──

[0042]  Table 10 ──────────────────────────────────── ── Day 3 Colony                                     Diameter (mm) Color ──────────────────────────────────── ── Candida albicans 3092 2.5 purple Candida albicans 3093 2.5 purple Candida albicans 3171 2.5 purple Candida albicans 3536 2.5 purple Candida tropicalis 3395 4.0 green Candida tropicalis 3172 3.5 green Candida krusei 3170 3.0 colorless Candida krusei 3389 3.0 colorless Candida glabrata 3388 2.5 colorless Candida glabrata 3390 2.0 colorless Candida parapsilosis 3091 1.5 colorless Candida parapsilosis 3169 1.5 colorless Candida guilliermondii 3426 1.5 colorless Candida famata 3777 2.0 colorless Trichosporon cutaneum 3455 2.0 colorless Trichosporon cutaneum 3456 2.0 colorless Trichosporon beigelii 3779 1.2 colorless Trichosporon beigelii 3780 1.2 colorless Esherichia coli 675 NG- Citrobacter freundii 389 NG- Proteus mirabilis 436 NG- Enterobacter cloacae 606 NG- Pseudomonas aeruginosa 1376 NG- ──────────────────────────────────── ──

[0043]

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) C12R 1:74) (C12Q 1/04 C12R 1: 725) (C12Q 1/04 C12R 1:74)

Claims (11)

[Claims] 1. Identification of Candida albicans, which comprises 5-bromo-6-chloro-3-indolyl-β-D-galactopyranoside (hereinafter abbreviated as magenta-gal) as a chromogenic enzyme substrate. Coloring medium for use. 2. The content of magenta-gal is 0.1-5 g / L.
The coloring medium for distinguishing Candida albicans according to claim 1, which is within the range. 3. The coloring medium for distinguishing Candida albicans according to claim 1 or 2, further comprising glucose. 4. The coloring medium for distinguishing Candida albicans according to claim 1, wherein the glucose content is in the range of 1 to 15 g / L. 5. Magenta-gal 0.1-5 per 1 L of culture medium
g, peptone 1 to 30 g, yeast extract 0.5 to 6.0
g, glucose 1 to 15 g, and agar 5 to 20 g, The color developing medium for distinguishing Candida albicans according to claim 1. 6. A color developing agent for distinguishing Candida tropicalis, which comprises 5-bromo-4-chloro-3-indolyl-β-D-glucopyranoside (hereinafter abbreviated as x-glc) as a chromogenic enzyme substrate. Culture medium. 7. The coloring medium for distinguishing Candida tropicalis according to claim 6, wherein the content of x-glc is in the range of 0.1 to 5 g / L. 8. The coloring medium for distinguishing Candida tropicalis according to claim 6 or 7, which further contains saccharose. 9. The coloring medium for distinguishing Candida tropicalis according to claim 6, wherein the sucrose content is in the range of 1 to 15 g / L. 10. x-glc 0.1-5 g per 1 L of medium,
The coloring medium for distinguishing Candida tropicalis according to claims 6 to 9, containing 1 to 30 g of peptone, 0.5 to 6.0 g of yeast extract, 1 to 15 g of sucrose, and 5 to 20 g of agar. 11. Magenta-gal and x as chromogenic enzyme substrates
A coloring medium for distinguishing Candida albicans and / or Candida tropicalis characterized by containing -glc.