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JP3298932B2 - Medium for selective culture of multidrug resistant staphylococci - Google Patents

Medium for selective culture of multidrug resistant staphylococci

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Publication number
JP3298932B2
JP3298932B2 JP18279992A JP18279992A JP3298932B2 JP 3298932 B2 JP3298932 B2 JP 3298932B2 JP 18279992 A JP18279992 A JP 18279992A JP 18279992 A JP18279992 A JP 18279992A JP 3298932 B2 JP3298932 B2 JP 3298932B2
Authority
JP
Japan
Prior art keywords
medium
staphylococci
antibiotic
present
selectively
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
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JP18279992A
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Japanese (ja)
Other versions
JPH07181A (en
Inventor
文章 田口
均 宮尾
華子 大久保
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Kyokuto Pharmaceutical Industrial Co Ltd
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Kyokuto Pharmaceutical Industrial Co Ltd
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Description

【発明の詳細な説明】DETAILED DESCRIPTION OF THE INVENTION

【0001】[0001]

【産業上の利用分野】本発明は、多剤耐性を示すブドウ
球菌、すなわちメチシリン耐性黄色ブドウ球菌(以下、
MRSAと略記する)および黄色ブドウ球菌以外のブドウ球
菌、例えばコアグラーゼ陰性ブドウ球菌(以下、CNS と
略記する)等のうち多剤耐性を示す菌を選択的に培養す
るための培地に関する。さらに詳しくは、ブドウ球菌感
染症の治療指針や、院内感染予防のための病院内環境汚
染状況の把握および改善に必要な情報を得るために有用
な上記培地に関する。
The present invention relates to staphylococci exhibiting multidrug resistance, that is, methicillin-resistant Staphylococcus aureus (hereinafter referred to as "methicillin-resistant Staphylococcus aureus").
The present invention relates to a medium for selectively culturing bacteria showing multidrug resistance among staphylococci other than Staphylococcus aureus, such as coagulase-negative staphylococci (hereinafter abbreviated as CNS). More particularly, the present invention relates to a medium useful for obtaining treatment guidelines for staphylococcal infections and obtaining information necessary for understanding and improving the status of environmental contamination in hospitals for preventing hospital-acquired infections.

【0002】[0002]

【従来の技術および発明が解決しようとする課題】患者
血液や尿などの検体中、食品中あるいは病院環境中のブ
ドウ球菌の検出には、従来使用されている細菌培地とし
て、スタフィロコックス培地No.110、マンニット
食塩加培地、食塩卵寒天基礎培地などがあり、これらの
培地はブドウ球菌を選択的に分離するための培地として
使用することができる。
BACKGROUND OF THE INVENTION For the detection of staphylococci in specimens such as patient blood and urine, in food or in a hospital environment, a staphylococcal medium No. 1 is used as a conventionally used bacterial medium. . 110, a mannitol supplemented medium, a salted egg agar base medium, and the like, and these mediums can be used as a medium for selectively separating staphylococci.

【0003】しかし、これらの培地はブドウ球菌を選択
的に増殖させることができるが、薬剤耐性ブドウ球菌で
あるか否かの鑑別をすることはできない。このため、薬
剤に対する耐性を知るには、これらのブドウ球菌選択培
地上にできたブドウ球菌の集落を採取し、集落の1つ1
つについて別個に薬剤感受性試験を行うことが必要であ
った(J. Clin. Microbiol.18:1084-1091,1983およびJ.
Clin. Microbiol.19:482-488,1984)。このように、従
来の方法では2段階の検査を行うため検査日数が長くな
り、また検査費用も高くなってしまう。
[0003] However, these media are capable of selectively growing staphylococci, but cannot identify whether they are drug-resistant staphylococci. For this reason, in order to know the resistance to the drug, the staphylococcal colonies formed on these staphylococcal selective media are collected, and each of the colonies is collected.
Separately, it was necessary to conduct a drug sensitivity test (J. Clin. Microbiol. 18: 1084-1091, 1983 and J. Clin.
Clin. Microbiol. 19: 482-488, 1984). As described above, in the conventional method, two-stage inspection is performed, so that the number of inspection days is increased and the inspection cost is increased.

【0004】さらにまた、上記のブドウ球菌選択培地
は、ブドウ球菌を選択的に分離培養するための培地であ
り、検体中の薬剤耐性の性質を保有するブドウ球菌の菌
数を知ることができないという問題もある。
Furthermore, the above-mentioned staphylococcal selective medium is a medium for selectively separating and culturing staphylococci, and it is said that the number of staphylococci having drug resistance in a specimen cannot be known. There are also problems.

【0005】そこで、メチシリン(感染症学雑誌、64
巻、5号、第549-556 頁)、セフチゾキシム(医学検
査、41巻、3号、第540 頁)等の抗生物質を単独で培地
に含有させて、抗生物質に耐性を示すブドウ球菌選択的
に増殖させる方法が試みられたが、多剤耐性のブドウ球
菌を選択的に増殖させる培地として実用可能なものは未
だ見出されていない。
Therefore, methicillin (Infectious Diseases Journal, 64
Vol. 5, No. 549-556) and antibiotics such as ceftizoxime (Medical Examination, Vol. 41, No. 3, pp. 540) alone in a medium to selectively show staphylococci resistant to antibiotics. However, no practical medium has been found as a medium for selectively growing multidrug-resistant staphylococci.

【0006】そこで本発明は、多剤耐性の性質を保有す
るブドウ球菌を効率的にかつ選択的に増殖させることの
できる培地を提供することを目的とする。
Accordingly, an object of the present invention is to provide a medium capable of efficiently and selectively growing staphylococci possessing multidrug resistance.

【0007】[0007]

【課題を解決するための手段】本発明者らは、多剤耐性
を示すブドウ球菌を選択的に分離培養するための培地に
ついて鋭意検討を重ねた結果、メチシリン耐性菌の指標
薬剤としてオキサシリン系抗生剤を用い、これと、見掛
上メチシリン感受性のブドウ球菌においてメチシリン耐
性を誘導することができる薬剤であり、しかも他のセフ
ェム系の薬剤と違って、不活化酵素により分解されにく
く、安定に培地中に存在することができるセフチゾキシ
ム系抗生剤とを組合せて用いると、多剤耐性を示すブド
ウ球菌を選択的に分離培養できることを見出した。
Means for Solving the Problems The present inventors have conducted intensive studies on a medium for selectively isolating and culturing staphylococci exhibiting multidrug resistance, and as a result, have found that oxacillin antibiotics can be used as an indicator for methicillin-resistant bacteria. It is a drug that can induce methicillin resistance in methicillin-sensitive staphylococci, and unlike other cephem-based drugs, it is not easily degraded by inactivating enzymes and stably It has been found that staphylococci exhibiting multidrug resistance can be selectively separated and cultured when used in combination with a ceftizoxime antibiotic which can be present in the bacterium.

【0008】すなわち本発明の培地は、ブドウ球菌を選
択的に培養するための培地に、多剤耐性を判定するため
の抗生物質として、培地1ml当たり、オキサシリン系
抗生剤を1.6μg以上およびセフチゾキシム系抗生剤
を6.25μg以上含むことを特徴とする。
[0008] That is, the medium of the present invention is used as an antibiotic for judging multidrug resistance in a medium for selectively cultivating staphylococci, at least 1.6 µg of oxacillin-based antibiotic and ceftizoxime per ml of medium. 6.25 μg or more of antibiotics.

【0009】「多剤耐性」とは一般に、メチシリン等の
β‐ラクタム系抗生剤を始めとし、現在広く使用されて
いる全ての抗生剤(例えばアミノグリコシド系、マクロ
ライド系、テトラサイクリン系等)のうちの複数薬剤に
対して耐性を示すことをいっている。
The term “multidrug resistance” generally refers to all antibiotics widely used at present (eg, aminoglycoside, macrolide, tetracycline, etc.), including β-lactam antibiotics such as methicillin. To be resistant to multiple drugs.

【0010】本発明の培地は、多剤耐性のブドウ球菌を
選択的に分離培養するために、培地中にオキサシリン系
抗生剤およびセフチゾキシム系抗生剤を組合せて含むこ
とを特徴とする。両者が含有されることが必須であり、
どちらか一方を含むだけでは本発明の効果を発揮できな
い。オキサシリン系抗生剤としては、6-アミノペニシラ
ン酸のメチルフェニルイソキサゾリル誘導体等を包含す
る。またセフチゾキシム系抗生剤としては、(6R,7R)-7-
[(Z)-2-(2-アミノ-4- チアゾリル)-2- メトキシイミノ
アセタミド]-8- オキソ-5- チア-1- アザビシクロ[4.
2.0]オクト-2-エン-2- カルボン酸、7-[(Z)-2-(2-アミ
ノ-4- チアゾリル)-2- メトキシイミノアセタミド]-3
- セフェム-4- カルボン酸等を包含する。オキサシリン
系抗生剤は培地1ml当たり、1.6μg以上の量で培
地中に含まれる。これより少ないと薬剤感受性菌の発育
を許すので、本発明の効果を発揮できない。含有量の上
限は特に必要ないが、多すぎると多剤耐性菌の発育抑制
を招くこともあるので、6.2μg以下が好ましい。使
用量の至適範囲は、培地1ml当たり1.6〜4.0μ
gである。また、セフチゾキシム系抗生剤は培地1ml
当たり6.25μg以上の量で培地中に含まれる。これ
より少ないと薬剤感受性菌の発育を許すので、本発明の
効果を発揮できない。含有量の上限は特に必要ないが、
多すぎると多剤耐性菌の発育抑制を招くこともあるの
で、25μg以下が好ましい。使用量の至適範囲は、培
地1ml当たり6.25〜12.5μgである。
[0010] The medium of the present invention is characterized in that the medium contains a combination of an oxacillin antibiotic and a ceftizoxime antibiotic in order to selectively isolate and culture multidrug-resistant staphylococci. It is essential that both are contained,
The effect of the present invention cannot be exhibited only by including one of them. Oxacillin antibiotics include methylphenyl isoxazolyl derivatives of 6-aminopenicillanic acid. As ceftizoxime antibiotics, (6R, 7R) -7-
[(Z) -2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamide] -8-oxo-5-thia-1-azabicyclo [4.
2.0] Oct-2-ene-2-carboxylic acid, 7-[(Z) -2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamide] -3
-Cephem-4-carboxylic acid and the like. The oxacillin antibiotic is contained in the medium in an amount of 1.6 μg or more per 1 ml of the medium. If the amount is less than this, the growth of drug-sensitive bacteria is allowed, so that the effects of the present invention cannot be exhibited. The upper limit of the content is not particularly required, but if it is too large, the growth of multidrug-resistant bacteria may be inhibited, so the content is preferably 6.2 μg or less. The optimal range of the amount used is 1.6 to 4.0 μm per 1 ml of the medium.
g. The ceftizoxime antibiotic is 1 ml of medium.
6.25 μg or more per medium. If the amount is less than this, the growth of drug-sensitive bacteria is allowed, so that the effects of the present invention cannot be exhibited. Although there is no particular upper limit on the content,
If the amount is too large, the growth of multidrug-resistant bacteria may be inhibited, so the amount is preferably 25 μg or less. The optimal range of the amount used is 6.25 to 12.5 μg per ml of the medium.

【0011】本発明の培地はまた、上記の2種の抗生剤
に加えてさらにゲンタマイシン系抗生剤を6.25μg
以上含むのが好ましい。これは、入院患者由来のMRSA株
の多くはゲンタマイシン耐性で、医療従事者より分離さ
れるMRSA株の多くはゲンタマイシン感受性であるという
ことから、多剤耐性菌のなかでも特に、ゲンタマイシン
耐性かどうかを判別することは臨床的意義が大きいから
である。使用するゲンタマイシン系抗生剤としては、ゲ
ンタマイシンC1 (C21435 7 )、ゲンタマイシ
ンC2 、ゲンタマイシンC1a等が包含される。ゲンタマ
イシン系抗生剤は、培地1ml当たり6.25μg以上
の量で培地中に含まれるのが好ましい。また、多すぎた
ときの多剤耐性菌の発育抑制を避けるためには25μg
以下が好ましい。使用量の至適範囲は、培地1ml当た
り6.25〜12.5μgである。
The medium of the present invention further contains 6.25 μg of a gentamicin antibiotic in addition to the above two antibiotics.
It is preferable to include the above. This is because many MRSA strains from hospitalized patients are gentamicin-resistant, and many MRSA strains isolated from healthcare workers are gentamicin-sensitive. Discrimination is of great clinical significance. Gentamicin antibiotics to be used include gentamicin C 1 (C 21 H 43 N 5 O 7 ), gentamicin C 2 , gentamicin C 1a and the like. The gentamicin antibiotic is preferably contained in the medium in an amount of 6.25 μg or more per ml of the medium. In addition, in order to avoid the suppression of the growth of multidrug-resistant bacteria when there is too much, 25 μg
The following is preferred. The optimal range of the amount used is 6.25 to 12.5 μg per ml of the medium.

【0012】本発明において、ブドウ球菌を選択的に培
養するための培地とは、当業者に公知の、ブドウ球菌培
養のための培地に慣用的に用いられる成分を含む培地を
いう。そのような成分としては例えば、カザミノ酸、ハ
ートエキス(ハートインフュージョン)、種々のペプト
ン、酵母エキス、肉エキス、水溶性デンプン、ブドウ
糖、L-トリプトファン、L-シスチン、ビオチン、CaC
2 、MgCl2 、NaCl、ピルビン酸(ナトリウ
ム)、卵黄液、寒天、マンニット、色素等が挙げられ
る。これらの成分を精製水に加えて培地を製造すること
ができる。
In the present invention, the medium for selectively cultivating staphylococci refers to a medium known to those skilled in the art and containing components commonly used in staphylococcal culture media. Such components include, for example, casamino acid, heart extract (heart infusion), various peptones, yeast extract, meat extract, water-soluble starch, glucose, L-tryptophan, L-cystine, biotin, CaC
l 2 , MgCl 2 , NaCl, pyruvate (sodium), egg yolk fluid, agar, mannitol, dyes and the like. These components can be added to purified water to produce a medium.

【0013】特に、ブドウ球菌のなかでもMRSAとその他
の多剤耐性ブドウ球菌(例えばCNS)を区別するために
は、卵黄反応とマンニット反応を組合せて使用するのが
好ましいので、培地に卵黄液、マンニットおよび色素を
含むのが好ましい。卵黄液は1〜10%(重量/培地体
積)含むのが好ましい。またマンニットは5〜20g/
培地1リットル、および色素(例えばブロムクレゾール
パープル、フェノールレッド、ニュートラルレッド、ブ
ロムチモールブルー等)は10〜30mg/培地1リッ
トル含むのが好ましい。卵黄反応により、MRSAは集落周
囲の黄濁環および、集落周囲培地表面の真珠光沢環を有
する集落として、その他は集落周囲を変化させない集落
として区別される。またマンニット反応により、MRSAは
集落周囲の培地色の黄変として、またその他は培地色を
変化させない集落として区別される。
Particularly, in order to distinguish MRSA from other staphylococci among other multidrug-resistant staphylococci (eg, CNS), it is preferable to use a combination of an egg yolk reaction and a mannitol reaction. , Mannitol and pigments. The yolk fluid preferably contains 1 to 10% (weight / medium volume). Mannit is 5 to 20 g /
It is preferable that 1 liter of the medium and the dye (for example, bromcresol purple, phenol red, neutral red, bromthymol blue, etc.) be contained in an amount of 10 to 30 mg / liter of the medium. By virtue of the yolk reaction, MRSA is distinguished as a colony having a cloudy ring around the colony and a pearlescent ring on the surface of the culture medium around the colony, and as a colony that does not change around the colony. Also, by the Mannit reaction, MRSA is distinguished as a yellowing of the medium color around the colony, and the other as a colony that does not change the medium color.

【0014】具体的なブドウ球菌培養用の培地組成の例
としては、例えばミュラーヒントン培地、トリプチケー
スソイ培地、ハートインフュージョン培地、普通寒天培
地等の基礎培地に、卵黄液、食塩、マンニット、ブドウ
糖、アミノ酸類、ピルビン酸等の付加的成分を添加した
ものが挙げられる。
Specific examples of the medium composition for staphylococcal culture include, for example, a basal medium such as Muller Hinton medium, trypticase soy medium, heart infusion medium, ordinary agar medium, egg yolk liquid, salt, mannitol , Glucose, amino acids, pyruvic acid and the like.

【0015】ブドウ球菌培養用培地の組成の好ましい例
としては、例えば次のような組成が挙げられる。すなわ
ち、精製水1000mlに、カザミノ酸14.8〜1
8.2g、ハートエキス2.7〜3.3g、水溶性デン
プン1.45〜1.65g、ブドウ糖5〜15g、L-ト
リプトファン0.045〜0.055g、L-シスチン
0.045〜0.055g、CaCl2 0.166〜
0.202g、MgCl20.188〜0.23g、N
aCl 6.8〜8.3g、ピルビン酸ナトリウム0〜
15g、卵黄液1〜10%(重量/体積)、寒天13.
5〜16.5g、マンニット5〜20gおよび色素10
〜30mgを添加する。
Preferred examples of the composition of the staphylococcal culture medium include the following compositions. That is, 14.8 to 1 casamino acids are added to 1000 ml of purified water.
8.2 g, heart extract 2.7-3.3 g, water-soluble starch 1.45-1.65 g, glucose 5-15 g, L-tryptophan 0.045-0.055 g, L-cystine 0.045-0.4 g. 055 g, CaCl 2 0.166-
0.202g, MgCl 2 0.188~0.23g, N
aCl 6.8-8.3 g, sodium pyruvate 0-
13. 15 g, egg yolk liquid 1-10% (weight / volume), agar
5 to 16.5 g, mannitol 5 to 20 g and pigment 10
Add 3030 mg.

【0016】本発明の培地のpHは、好ましくは7.0
〜7.8、至適pHは7.4±0.2である。
The pH of the medium of the present invention is preferably 7.0.
77.8, the optimum pH is 7.4 ± 0.2.

【0017】本発明の培地は、常法により培養を行うこ
とができる。例えば菌体を塗抹した培地を、通常32〜
38℃、特に好ましくは35℃前後で、通常18〜48
時間、好ましくは約24時間培養を行う。
The medium of the present invention can be cultured by a conventional method. For example, a medium in which cells are smeared is usually 32 to
38 ° C, particularly preferably around 35 ° C, usually 18 to 48
The culture is performed for a time, preferably for about 24 hours.

【0018】[0018]

【実施例】以下の実施例により本発明をさらに詳しく説
明する。なお、実施例では、次の培地を使用した。本発明の平板培地A オキサシリン 4mg、セフチゾキシム 12.5m
g、カザミノ酸 15.5g、ハートエキス 3.0
g、水溶性デンプン 1.5g、ブドウ糖 2.0g、
L-トリプトファン 0.05g、L-シスチン 0.05
g、CaCl2 0.15g、MgCl2 0.1g、
NaCl 75g、ピルビン酸ナトリウム1g、寒天
15g、マンニット 10g、およびブロムクレゾール
パープル20mgに精製水1000mlを加えた。これ
を滅菌した後、卵黄液を5%(重量/体積)の割合に加
え、シャーレに各20ml流し入れ、固化させた。本発明の平板培地B オキサシリン 4mg、セフチゾキシム 12.5m
g、ゲンタマイシン 12.5mg、カザミノ酸 1
5.5g、ハートエキス 3.0g、水溶性デンプン
1.5g、ブドウ糖 2.0g、L-トリプトファン
0.05g、L-シスチン 0.05g、CaCl2
0.15g、MgCl2 0.1g、NaCl75g、
ピルビン酸ナトリウム 1g、寒天 15g、マンニッ
ト 10g、およびブロムクレゾールパープル 20m
gに精製水1000mlを加えた。これを滅菌した後、
卵黄液を5%(重量/体積)の割合に加え、シャーレに
各20ml流し入れ、固化させた。比較例の平板培地A (抗生物質を含まないこと以外は本
発明の培地と等しい成分を含有) カザミノ酸 15.5g、ハートエキス 3.0g、水
溶性デンプン 1.5g、ブドウ糖 2.0g、L-トリ
プトファン 0.05g、L-シスチン 0.05g、C
aCl2 0.15g、MgCl2 0.1g、NaC
l 75g、ピルビン酸ナトリウム 1g、寒天 15
g、マンニット 10g、およびブロムクレゾールパー
プル 20mgに精製水1000mlを加えた。これを
滅菌した後、卵黄液を5%(重量/体積)の割合に加
え、シャーレに各20ml流し入れ、固化させた。比較例の平板培地B(マンニット食塩卵黄培地) 肉エキス 1.0g、ペプトン 10.0g、マンニッ
ト 10.0g、NaCl 75.0g、フェノールレ
ッド 0.025gおよび寒天 15.0g(極東マン
ニットソルト寒天培地として極東製薬工業株式会社より
市販)に精製水1000mlを加えた。これを滅菌した
後、卵黄液を5%(重量/体積)の割合に加え、シャー
レに各20ml流し入れ、固化させた。実施例1〜2および比較例1〜2 MRSAと判明している黄色ブドウ球菌20株およびメチシ
リン感受性黄色ブドウ球菌(以下、MSSAと略記する)2
0株を、本発明の平板培地、比較例の平板培地Aおよび
比較例の平板培地Bのそれぞれに塗抹し、35℃で約2
4時間培養を行い、黄色ブドウ球菌の増殖を検討した。
結果を表1に示す。
The present invention will be described in more detail with reference to the following examples. In the examples, the following media were used. Plate medium A of the present invention A oxacillin 4 mg, ceftizoxime 12.5 m
g, casamino acid 15.5 g, heart extract 3.0
g, water-soluble starch 1.5 g, glucose 2.0 g,
L-tryptophan 0.05g, L-cystine 0.05
g, CaCl 2 0.15 g, MgCl 2 0.1 g,
NaCl 75 g, sodium pyruvate 1 g, agar
To 15 g, 10 g of mannitol, and 20 mg of bromcresol purple, 1000 ml of purified water was added. After sterilization, the yolk liquid was added to a ratio of 5% (weight / volume), and 20 ml of each was poured into a petri dish and solidified. Plate medium B of the present invention B Oxacillin 4 mg, Ceftizoxime 12.5 m
g, gentamicin 12.5 mg, casamino acid 1
5.5 g, heart extract 3.0 g, water-soluble starch
1.5g, glucose 2.0g, L-tryptophan
0.05 g, L-cystine 0.05 g, CaCl 2
0.15 g, MgCl 2 0.1 g, NaCl 75 g,
Sodium pyruvate 1 g, agar 15 g, mannitol 10 g, and bromcresol purple 20 m
g of purified water was added to the mixture. After sterilizing this,
Egg yolk liquid was added to a ratio of 5% (weight / volume), and 20 ml of each was poured into a petri dish and solidified. Plate medium A of Comparative Example (containing the same components as the medium of the present invention except that it does not contain antibiotics) 15.5 g of casamino acid, 3.0 g of heart extract, 1.5 g of water-soluble starch, 2.0 g of glucose, L -Tryptophan 0.05g, L-cystine 0.05g, C
aCl 2 0.15 g, MgCl 2 0.1 g, NaC
l 75 g, sodium pyruvate 1 g, agar 15
g, mannitol 10 g, and bromcresol purple 20 mg, purified water 1000 ml was added. After sterilization, the yolk liquid was added to a ratio of 5% (weight / volume), and 20 ml of each was poured into a petri dish and solidified. Plate Medium B (Mannit Salt Egg Yolk Medium) Meat Extract 1.0 g, Peptone 10.0 g, Mannit 10.0 g, NaCl 75.0 g, Phenol Red 0.025 g, and Agar 15.0 g (Far East Mannit Salt) 1000 ml of purified water was added to an agar medium (commercially available from Kyokuto Pharmaceutical Co., Ltd.). After sterilization, the yolk liquid was added to a ratio of 5% (weight / volume), and 20 ml of each was poured into a petri dish and solidified. Examples 1-2 and Comparative Examples 1-2 Twenty strains of S. aureus known as MRSA and methicillin-sensitive S. aureus (hereinafter abbreviated as MSSA) 2
0 strains were spread on each of the plate medium of the present invention, the plate medium A of the comparative example, and the plate medium B of the comparative example.
After culturing for 4 hours, the growth of Staphylococcus aureus was examined.
Table 1 shows the results.

【0019】[0019]

【表1】 [Table 1]

【0020】比較例の平板培地においてはいずれも、試
験菌株すべてが増殖しているが、本発明の平板培地にお
いては、MRSAのみが選択的に増殖した。実施例3および比較例3〜4 多剤耐性CNS であると判明しているCNS 20株およびメ
チシリン感受性のCNS20株を、本発明の平板培地、比
較例平板培地Aおよび比較例平板培地Bのそれぞれに塗
抹し、35℃で約24時間培養を行い、CNS の増殖を検
討した。結果を表2に示す。
All the test strains grew in the plate medium of the comparative example, but only the MRSA grew selectively in the plate medium of the present invention. Example 3 and Comparative Examples 3 and 4 20 CNS strains known to be multidrug-resistant CNS and 20 methicillin-sensitive CNS strains were used for the plate medium of the present invention, the comparative plate medium A and the comparative plate medium B, respectively. And cultured at 35 ° C. for about 24 hours to examine the proliferation of CNS. Table 2 shows the results.

【0021】[0021]

【表2】 [Table 2]

【0022】比較例の平板培地においてはいずれも、試
験菌株すべてが増殖しているが、本発明の平板培地にお
いては、多剤耐性を示すメチシリン耐性CNS のみが選択
的に増殖した。実施例4 病院従事者間におけるMRSA保有状況を把握するため、病
院従事者5人(A,B,CおよびD)の鼻前底、咽頭お
よび手のひらの皮膚表面を滅菌綿棒にて拭い、この綿棒
を1mlの滅菌生理食塩水に良くほぐしたものを検体と
して使用した。この検体0.1mlを本発明の平板培地
表面に均一に塗抹し、37℃で約24時間培養を行っ
て、菌の増殖を検討した。なお、MRSA以外のブドウ球菌
の集落については、家兎プラズマ凝固試験(コアグラー
ゼ試験)においてすべての株が陰性であることにより、
多剤耐性CNS であることを判定した。結果を表3に示
す。
All the test strains grew in the plate medium of the comparative example, but only the methicillin-resistant CNS showing multidrug resistance grew selectively in the plate medium of the present invention. Example 4 In order to grasp the MRSA possession status among hospital workers, the skin surfaces of the nasal base, pharynx and palms of five hospital workers (A, B, C and D) were wiped with a sterile cotton swab. Was thoroughly dissolved in 1 ml of sterile physiological saline and used as a sample. 0.1 ml of this sample was uniformly spread on the surface of the plate medium of the present invention, and cultured at 37 ° C. for about 24 hours to examine the growth of bacteria. Regarding staphylococcal colonies other than MRSA, all strains were negative in the rabbit plasma coagulation test (coagulase test).
It was determined to be a multidrug resistant CNS. Table 3 shows the results.

【0023】[0023]

【表3】 [Table 3]

【0024】[0024]

【発明の効果】本発明によれば、多剤耐性を示すブドウ
球菌を選択的に増殖させて検出することができる。よっ
て、本発明の培地は非常に有用である。
According to the present invention, staphylococci exhibiting multidrug resistance can be selectively grown and detected. Therefore, the medium of the present invention is very useful.

───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 平5−227992(JP,A) (58)調査した分野(Int.Cl.7,DB名) C12N 1/20 - 1/38 C12Q 1/00 - 1/14 BIOSIS(DIALOG) JICSTファイル(JOIS)────────────────────────────────────────────────── (5) References JP-A-5-227992 (JP, A) (58) Fields investigated (Int. Cl. 7 , DB name) C12N 1/20-1/38 C12Q 1 / 00-1/14 BIOSIS (DIALOG) JICST file (JOIS)

Claims (3)

(57)【特許請求の範囲】(57) [Claims] 【請求項1】 ブドウ球菌を選択的に培養するための培
地に、多剤耐性を判定するための抗生物質として、培地
1ml当たり、オキサシリン系抗生剤を1.6μg以上
およびセフチゾキシム系抗生剤を6.25μg以上含む
ことを特徴とする多剤耐性ブドウ球菌培養用培地。
1. As an antibiotic for judging multidrug resistance in a medium for selectively culturing staphylococci, 1.6 μg or more of an oxacillin antibiotic and 6 or more of a ceftizoxime antibiotic are used per 1 ml of the medium. A multi-drug resistant staphylococcal culture medium comprising at least 25 μg.
【請求項2】 さらに、培地1ml当たり、ゲンタマイ
シン系抗生剤を6.25μg以上含む請求項1記載の培
地。
2. The medium according to claim 1, further comprising 6.25 μg or more of a gentamicin antibiotic per 1 ml of the medium.
【請求項3】 ブドウ球菌を選択的に培養するための培
地が、精製水1000mlに、カザミノ酸14.8〜1
8.2g、ハートエキス2.7〜3.3g、水溶性デン
プン1.45〜1.65g、ブドウ糖5〜15g、L-ト
リプトファン0.045〜0.055g、L-シスチン
0.045〜0.055g、CaCl20.166〜
0.202g、MgCl2 0.188〜0.23g、
NaCl6.8〜8.3g、ピルビン酸ナトリウム0〜
15g、卵黄液1〜10%(重量/体積)、寒天13.
5〜16.5g、マンニット5〜20gおよび色素10
〜30mgを添加してなるものである請求項1または2
項に記載の培地。
3. A culture medium for selectively culturing staphylococci is prepared by adding casamino acids 14.8 to 1 to 1,000 ml of purified water.
8.2 g, heart extract 2.7-3.3 g, water-soluble starch 1.45-1.65 g, glucose 5-15 g, L-tryptophan 0.045-0.055 g, L-cystine 0.045-0.4 g. 055 g, CaCl 2 0.166-
0.202g, MgCl 2 0.188~0.23g,
6.8-8.3 g of NaCl, 0-sodium pyruvate
13. 15 g, egg yolk liquid 1-10% (weight / volume), agar
5 to 16.5 g, mannitol 5 to 20 g and pigment 10
3. The composition according to claim 1 or 2, wherein the amount is from 30 to 30 mg.
The medium according to Item.
JP18279992A 1992-06-18 1992-06-18 Medium for selective culture of multidrug resistant staphylococci Expired - Lifetime JP3298932B2 (en)

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JPH07181A JPH07181A (en) 1995-01-06
JP3298932B2 true JP3298932B2 (en) 2002-07-08

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FR2844807B1 (en) * 2002-09-23 2005-11-11 Rambach Alain METHOD FOR DETECTING MICROORGANISMS RESISTANT TO METICILLIN
JP2006515184A (en) * 2003-01-10 2006-05-25 ベクトン・ディキンソン・アンド・カンパニー Methods for detecting antibiotic resistance in microorganisms
FR2937052A1 (en) 2008-10-08 2010-04-16 Biomerieux Sa REACTIONAL MEDIUM FOR STAPHYLOCOCCUS AUREUS BACTERIA
FR2936816B1 (en) * 2008-10-08 2013-03-22 Biomerieux Sa REACTION ENVIRONMENT FOR STAPHYLOCOCUS AUREUS-RESISTANT BACTERIA (MRSA)
JP5515139B2 (en) * 2008-10-21 2014-06-11 国立大学法人名古屋大学 Multidrug-resistant Pseudomonas aeruginosa screening medium
FR2949119B1 (en) * 2009-08-13 2011-08-26 Biomerieux Sa REACTION ENVIRONMENT FOR STAPHYLOCOCUS AUREUS-RESISTANT BACTERIA (MRSA)
US8497086B2 (en) 2009-08-13 2013-07-30 Biomereux Reaction medium for methicillin-resistant Staphylococcus aureus (MRSA) bacteria
US20110275114A1 (en) * 2010-05-06 2011-11-10 Edberg Stephen C Method and medium for detecting the presence or absence of methicillin resistant staphylococcus aureus (mrsa) in a test sample
JP2015126756A (en) * 2015-04-10 2015-07-09 栄研化学株式会社 Potentiation of color development reaction and/or fluorescent color development reaction with egg yolk liquid

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