JPH0556960B2 - - Google Patents
Info
- Publication number
- JPH0556960B2 JPH0556960B2 JP28917785A JP28917785A JPH0556960B2 JP H0556960 B2 JPH0556960 B2 JP H0556960B2 JP 28917785 A JP28917785 A JP 28917785A JP 28917785 A JP28917785 A JP 28917785A JP H0556960 B2 JPH0556960 B2 JP H0556960B2
- Authority
- JP
- Japan
- Prior art keywords
- starch
- bacteria
- medium
- metal salt
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 229920002472 Starch Polymers 0.000 claims description 25
- 239000008107 starch Substances 0.000 claims description 25
- 235000019698 starch Nutrition 0.000 claims description 24
- 238000012360 testing method Methods 0.000 claims description 23
- 239000013028 medium composition Substances 0.000 claims description 11
- 239000001963 growth medium Substances 0.000 claims description 10
- 229910052751 metal Inorganic materials 0.000 claims description 8
- 239000002184 metal Substances 0.000 claims description 8
- 150000003839 salts Chemical class 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- 244000052616 bacterial pathogen Species 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 claims description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-M dihydrogenphosphate Chemical compound OP(O)([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-M 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical group [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical group [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims 1
- 241000894006 Bacteria Species 0.000 description 18
- 239000002609 medium Substances 0.000 description 11
- 229920001817 Agar Polymers 0.000 description 8
- 239000008272 agar Substances 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000001802 infusion Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 238000007796 conventional method Methods 0.000 description 3
- 208000035143 Bacterial infection Diseases 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000589565 Flavobacterium Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 229910001514 alkali metal chloride Inorganic materials 0.000 description 1
- -1 alkali metal salt Chemical class 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
【発明の詳細な説明】
発明の背景
技術分野
本発明は微生物のデンプン利用能検査用培地組
成物に関する。さらに詳しくは、本発明はグルコ
ース非発酵性グラム陰性桿菌のデンプン利用能検
査用培地組成物に関するものである。
細菌感染症に対して適切な治療を施すためには
病原菌を同定し、感受性試験を行い、その病原菌
に有効な薬剤を決定することが重要である。この
ような病原菌の同定に際しては多項目にわたる生
化学的性状検査が行われ、その項目の１つとして
グルコース非発酵性グラム陰性桿菌についてデン
プン利用能の検査が行われる。これは、上記菌が
有するデンプン加水分解能が利用し、菌の同定を
行うものである。本発明の培地組成物はこのよう
な試験に好適に使用される。
先行技術およびその問題点
上記検査の培地として従来、普通寒天培地、
SCD（ソイビーンカゼインダイジエスト）寒天培
地等に0.1〜0.2％のデンプンを加えた培地、ある
いはミユーラーヒントン寒天培地（0.15％のデン
プンを含有）に菌を接種し、30℃で１〜２日間培
養後、菌の発育している部分にルゴール液を加え
て判定している。デンプンが加水分解されていれ
ばその部分は透明となり、加水分解されていない
場合は黒褐色になる。このように従来培地におい
てもデンプン利用能の検査は可能であるが、培地
素材に寒天を使用しているため反応速度が遅く、
判定に１〜２日を要する。
細菌感染症の早期治療のためには、菌の同定は
できるだけ速やかに行うことが必要であり、より
短かい時間で菌の同定が可能な培地の出現が望ま
れていた。しかし、一方においては、検査作業の
都合上、判定を翌日に行なわざるを得ない場合も
生じ、このような場合には培養期間が厳格に規制
されておらず、都合により培養時間を延長しても
反応過剰になつたりせず、正確な判定が可能な培
地が望ましい。
発明の目的
本発明は短時間の培養で同定が可能であるとと
もに長時間培養しても正確な同定が可能であるグ
ルコース非発酵性グラム陰性桿菌のデンプン利用
能の検査用培地組成物を提供することを目的とす
る。
本発明はさらに、呈色反応が明瞭であり、判定
が容易な上記検査用培地組成物を提供することを
目的とする。
上記の本発明の目的はデンプン1.0部（重量部、
以下同じ）に対し、ペプトン0.4〜4.0部、リン酸
二水素一アルカリ金属塩0.02〜0.2部、およびリ
ン酸一水素二アルカリ金属塩0.18〜1.8部の組成
よりなるグルコース非発酵性グラム陰性桿菌のデ
ンプン利用能検査用培地組成物によつて達成され
る。
発明の具体的説明
本発明の培地組成物において、デンプンは基質
であり、これが検査菌によつて加水分解されると
培地のヨー素デンプン反応の色が薄くなる。この
変化によつて検査菌のデンプン利用能を判定す
る。ペプトンは養分として用いられる。リン酸二
水素一アルカリ金属塩とリン酸一水素二アルカリ
金属塩はPH調整剤であり、菌の発育に最適なPH
7.0〜7.6になるような範囲に設定されている。本
発明の培地組成物にはさらに浸透圧調整剤として
アルカリ金属塩２〜30部が使用されるのが望まし
く、塩化ナトリウム、塩化カリウムのようなアル
カリ金属塩化物が好適である。
本発明の培地組成物の前述した成分割合は臨界
的であり、特にデンプン濃度は重要である。デン
プン濃度が低すぎると偽陽性の判定となりやす
く、逆に高すぎると偽陰性の判定となりやすい。
本発明の培地では、上述した偽陽性や偽陰性の誤
まつた判定を下す危険が少なく、かつ、接種菌数
を適宜選択することにより短時間培養（４〜５時
間）または一昼夜培養（18〜22時間）のいずれに
よつても正しい判定が可能なようにデンプン濃度
が設定されている。
本発明の培地組成物は常法に従つて所定量の各
成分を水に溶解して使用される。近年生化学的性
状検査は多数の試験を多穴プレート上で同時に行
うのが普通であり、このような場合には本発明培
地を試験用プレートのウエルに注入し、乾燥させ
て乾燥培地とする。試験に際しては該ウエルに試
験菌の懸濁水を所定量分注し、30℃で所定時間培
養する。４〜５時間の培養で判定することが望ま
れる場合は、液体培地50μ当り約7.5×107個の
菌を接種し、18〜22時間培養後の判定が望まれる
場合は約1.5×107個の菌を接種するのが好まし
い。このように接種菌数を調整することにより、
短時間でもまた翌日でも検査が行える。
次に本発明の培地組成物を使用して細菌のデン
プン利用能を検査した実施例を示す。
実施例
表１に記載の組成を有する本発明培地組成物を
常法に従つて蒸留水１に溶解し、菌同定用培地
１〜３を得た。DETAILED DESCRIPTION OF THE INVENTION BACKGROUND OF THE INVENTION Technical Field The present invention relates to a medium composition for testing the starch utilization ability of microorganisms. More specifically, the present invention relates to a medium composition for testing the starch utilization ability of glucose non-fermenting Gram-negative bacilli. In order to provide appropriate treatment for bacterial infections, it is important to identify the pathogen, conduct susceptibility tests, and determine effective drugs against the pathogen. When identifying such pathogenic bacteria, a multi-item biochemical property test is performed, and one of the items is a test for starch utilization ability of glucose non-fermenting Gram-negative rods. This utilizes the starch hydrolyzing ability of the above-mentioned bacteria to identify the bacteria. The medium composition of the present invention is suitably used for such tests. Prior art and its problems Traditionally, ordinary agar medium,
Bacteria are inoculated onto SCD (soybean casein digest) agar medium containing 0.1-0.2% starch, or Mueller-Hinton agar medium (containing 0.15% starch), and cultured at 30°C for 1-2 days. Afterwards, Lugol's solution is added to the area where the bacteria are growing to make a determination. If the starch is hydrolyzed, it will be transparent; if it is not, it will be dark brown. Although it is possible to test starch utilization using conventional media, the reaction rate is slow because agar is used as the media material.
It takes 1 to 2 days for the determination. For early treatment of bacterial infections, it is necessary to identify bacteria as quickly as possible, and there has been a desire for a culture medium that can identify bacteria in a shorter time. However, on the other hand, due to the convenience of testing work, there are cases where the judgment has to be made the next day, and in such cases, the culture period is not strictly regulated and the culture time may be extended for convenience. It is desirable to use a medium that does not overreact and allows accurate determination. Purpose of the Invention The present invention provides a medium composition for testing the starch utilization ability of glucose non-fermenting Gram-negative bacilli, which can be identified in a short time culture and can be accurately identified even in a long time culture. The purpose is to A further object of the present invention is to provide the above-mentioned test medium composition that has a clear color reaction and is easy to judge. The above object of the present invention is to use 1.0 parts of starch (parts by weight,
The same applies hereinafter) to 0.4 to 4.0 parts of peptone, 0.02 to 0.2 parts of a monoalkali metal salt of dihydrogen phosphate, and 0.18 to 1.8 parts of a dialkali metal salt of monohydrogen phosphate. This is achieved by a medium composition for testing starch availability. DETAILED DESCRIPTION OF THE INVENTION In the culture medium composition of the present invention, starch is the substrate, and when it is hydrolyzed by the test bacteria, the color of the iodine-starch reaction in the culture medium becomes lighter. Based on this change, the ability of the test bacteria to utilize starch is determined. Peptone is used as a nutrient. Monoalkali metal dihydrogen phosphate and dialkali metal monohydrogen phosphate are PH regulators, and the pH is optimal for bacterial growth.
It is set to a range of 7.0 to 7.6. It is desirable that the medium composition of the present invention further contains 2 to 30 parts of an alkali metal salt as an osmotic pressure regulator, with alkali metal chlorides such as sodium chloride and potassium chloride being preferred. The above-mentioned component ratios of the culture medium composition of the present invention are critical, and the starch concentration is particularly important. If the starch concentration is too low, a false positive determination is likely to occur; conversely, if it is too high, a false negative determination is likely to occur.
The culture medium of the present invention has a low risk of making false positive or false negative judgments as described above, and can be cultured for a short time (4 to 5 hours) or overnight (18 to 5 hours) by appropriately selecting the number of inoculated bacteria. The starch concentration is set so that correct judgment can be made regardless of the time (22 hours). The medium composition of the present invention is used by dissolving predetermined amounts of each component in water according to a conventional method. In recent years, it has become common for biochemical property tests to simultaneously perform multiple tests on multi-well plates, and in such cases, the culture medium of the present invention is injected into the wells of the test plate and dried to form a dry medium. . During the test, a predetermined amount of suspension water of test bacteria is dispensed into the well, and cultured at 30°C for a predetermined time. If it is desired to make a judgment after culturing for 4 to 5 hours, inoculate about 7.5 x 107 bacteria per 50μ of liquid medium, and if you want to make a judgment after culturing for 18 to 22 hours, about 1.5 x 107 bacteria. It is preferable to inoculate the bacteria. By adjusting the number of inoculated bacteria in this way,
Tests can be performed in a short period of time or even the next day. Next, an example will be shown in which the ability of bacteria to utilize starch was tested using the medium composition of the present invention. Example The culture medium composition of the present invention having the composition shown in Table 1 was dissolved in distilled water 1 according to a conventional method to obtain culture media 1 to 3 for bacterial identification.
【表】
上記培地１〜３の多穴プレートの各ウエルに
50μずつ分注し、40℃で乾燥した。次に寒天培
地上で各種細菌を30℃で18〜24時間培養し、1.0
mlの減菌蒸留水注に培養時間が４時間の場合は
1.5×109個／mlまた20時間の場合３×108個／ml
となるように懸濁し各ウエルに50μずつ接種
し、30℃で所定時間培養した。比較試験として、
下記のハートインフユージヨン寒天（デンプン含
有）培地を用いた従来法により培養を行なつた。
ルゴール液によるヨー素デンプン反応によりデン
プン利用能の有無を判定した。結果を表２に示
す。
ハートインフユージヨン寒天培地組成
ウシ心筋浸出液（心筋500ｇからの浸出液）
1000ml
ペプトン 10ｇ
NaCl ５ｇ
寒 天 15ｇ
デンプン ２ｇ
PH7.4[Table] In each well of the multi-well plate for the above media 1 to 3.
It was dispensed into 50μ portions and dried at 40°C. Next, various bacteria were cultured on agar medium at 30°C for 18 to 24 hours, and 1.0
If the incubation time is 4 hours in ml of sterile distilled water,
1.5×10 9 pieces/ml or 3×10 8 pieces/ml for 20 hours
The cells were suspended in such a manner that 50μ of each well was inoculated, and cultured at 30°C for a predetermined period of time. As a comparative test,
Culture was carried out by a conventional method using the heart infusion agar medium (containing starch) described below.
The presence or absence of starch utilization ability was determined by iodine starch reaction using Lugol's solution. The results are shown in Table 2. Heart Infusion Agar Medium Composition Bovine myocardial infusion (infusion from 500g of myocardium)
1000ml Peptone 10g NaCl 5g Agar 15g Starch 2g PH7.4
【表】
発明の具体的作用および効果
表２から明らかなように、グルコース非発酵性
グラム陰性桿菌のデンプン利用能検査において、
本発明の培地を使用すると、培養時間の長短を問
わず正しい菌の同定が可能である。従つて検査作
業の都合により即日同定、翌日同定のいずれも選
択することができる。これに対して対照培地は少
くとも20時間の培養が必要であり、即日判定は不
可能である。例えば、対照培地を使用した場合
は、シユードモナス・スタツチエリATCC17588、
およびフラボバクテリウム・インドロゲネス
ATCC29897は４時間培養では反応は陰性であり
20時間培養で陽性となる。これに対して本発明の
培地を使用すると、いずれの菌においても４時間
培養で正しい判定が可能である。[Table] Specific functions and effects of the invention As is clear from Table 2, in the starch utilization ability test of glucose non-fermenting Gram-negative bacilli,
By using the culture medium of the present invention, it is possible to correctly identify bacteria regardless of the length of the culture time. Therefore, either same-day identification or next-day identification can be selected depending on the convenience of the inspection work. On the other hand, the control medium requires cultivation for at least 20 hours, and same-day determination is not possible. For example, if control medium was used, Pseudomonas stutschieri ATCC17588,
and Flavobacterium indrogenes
ATCC29897 showed a negative reaction when cultured for 4 hours.
Test becomes positive after 20 hours of culture. On the other hand, when the medium of the present invention is used, correct determination can be made for any bacteria after 4 hours of culture.
Claims (1)
ペプトン0.4〜4.0部、リン酸二水素一アルカリ金
属塩0.02〜0.2部、およびリン酸一水素二アルカ
リ金属塩0.18〜1.8部の組成よりなるグルコース
非発酵性グラム陰性桿菌のデンプン利用能検査用
培地組成物。 ２ リン酸二水素一アルカリ金属塩がリン酸二水
素一カリウム塩であり、リン酸一水素二アルカリ
金属塩がリン酸一水素二ナトリウム塩である特許
請求の範囲第１項記載の培地組成物。[Claims] 1. For 1.0 part of starch (part by weight, same hereinafter),
Culture medium for testing starch utilization of glucose non-fermenting Gram-negative bacilli, consisting of 0.4 to 4.0 parts of peptone, 0.02 to 0.2 parts of a monoalkali metal salt of dihydrogen phosphate, and 0.18 to 1.8 parts of a dialkali metal salt of monohydrogen phosphate. Composition. 2. The medium composition according to claim 1, wherein the monoalkali metal salt of dihydrogen phosphate is monopotassium dihydrogen phosphate, and the dialkali metal salt of monohydrogen phosphate is disodium monohydrogen phosphate. .
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP28917785A JPS62151197A (en) | 1985-12-24 | 1985-12-24 | Culture medium composition for testing starch utilization ability |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP28917785A JPS62151197A (en) | 1985-12-24 | 1985-12-24 | Culture medium composition for testing starch utilization ability |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS62151197A JPS62151197A (en) | 1987-07-06 |
| JPH0556960B2 true JPH0556960B2 (en) | 1993-08-20 |
Family
ID=17739763
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP28917785A Granted JPS62151197A (en) | 1985-12-24 | 1985-12-24 | Culture medium composition for testing starch utilization ability |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS62151197A (en) |
-
1985
- 1985-12-24 JP JP28917785A patent/JPS62151197A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS62151197A (en) | 1987-07-06 |
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