JP4414176B2 - カンジダ鑑別用発色培地 - Google Patents
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- JP4414176B2 JP4414176B2 JP2003316533A JP2003316533A JP4414176B2 JP 4414176 B2 JP4414176 B2 JP 4414176B2 JP 2003316533 A JP2003316533 A JP 2003316533A JP 2003316533 A JP2003316533 A JP 2003316533A JP 4414176 B2 JP4414176 B2 JP 4414176B2
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
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Description
このような菌数の確認については、培養法が汎用されており、既に水野−高田培地やサブロー培地などの種々の培地が市販されている。これらの培地は、いずれも炭素源、窒素源の他、一般細菌の生育を抑制する物質も含みカンジダ菌のみの検出を目的としている
(例えば、非特許文献1、非特許文献2参照)。
しかしながら、これらの培地では菌の培養に2〜3日も要するために速やかな診断を下すことができず、迅速な治療開始ができないという問題があった。
従って本発明は、このような従来の課題に着目してなされたものであって、5種類のカンジダ菌を20時間程度の培養で鑑別することのできるカンジダ鑑別用発色培地を提供することを目的とする。
(1)酵素基質および酸化還元試薬を含有することを特徴とするカンジダ鑑別用発色培地。
(2)酵素基質がx-phosおよび/またはx-galactosaminideである(1)記載のカンジダ鑑別用発色培地。
(3)酵素基質の含有量が0.01〜0.1g/Lの範囲である(1)または(2)記載のカンジダ鑑別用発色培地。
(4)酸化還元試薬がテトラゾリウム塩である(1)記載のカンジダ鑑別用発色培地。
(5)テトラゾリウム塩がTTVおよび/またはTTBである(4)記載のカンジダ鑑別用発色培地。
(6)酸化還元試薬の含有量が0.001〜0.01g/Lの範囲である(1)、(4)または(5)記載のカンジダ鑑別用発色培地。
(7)培地1Lあたり、ペプトン10〜30g、酵母エキス1〜10g、白糖1〜20g、ブドウ糖1〜10g、TTV0.001〜0.01g、x-phos0.01〜0.1gおよび寒天3〜20gを含有する(1)〜(6)記載のカンジダ鑑別用発色培地。
本発明のカンジダ鑑別用発色培地の特徴は、培地中に酵素基質および酸化還元試薬を含有させることにある。
本発明においては、酵素基質と酸化還元試薬とが相俟って相乗的効果を生じ、5種類のカンジダ菌が短時間でそれぞれ特有の色を持つコロニーを形成するため容易に鑑別することができる。即ち、培養20時間程度で、カンジダ・アルビカンス(C.albicans)は桃色、カンジダ・グラブラータ(C.glabrata)は白色、カンジダ・トロピカリス(C.tropicalis)は青色、カンジダ・クルセイ(C.krusei)は水色、カンジダ・パラプシロシス(C.parapsilsis)は濃桃色にそれぞれ発色する。
これらの成分の他に、カンジダ菌の発育を促進する成分としては、硫酸アンモニウムなどの窒素源、マルトースやスクロースなどの糖類、さらに必要に応じてミネラルやビタミンなどの任意のものを含めることができる。
固形培地の固化剤としては、寒天、カラギーナン、ローカストビーンガムなど通常使用されているものが挙げられる。
表1に示した培地成分のうちTTVを除いた51.35gを秤量し、1000mLの精製水に溶解した。溶解後、pHを6.0±0.2に調整し、100℃で30分間加熱溶解した。50℃に冷却後、0.006gのTTVを少量の蒸留水に溶解し、濾過滅菌して加えた後、20mLづつシャーレに分注して固化した。固化後、30分間乾燥した。対照としてクロモカンジダ培地を作成した。
表1 培地の処方 g/L
──────────────────────────
カゼインペプトン 15.0
酵母エキス 5.0
白糖 10.0
ブドウ糖 5.0
クロラムフェニコール 0.3
TTV 0.006
X−phos.2Na 0.05
寒天 7.0
カラギーナン(カッパータイプ) 3. 5
カラギーナン(イオタタイプ) 2.3
ローカストビーンガム 3.2
──────────────────────────
pH6.0±0.2
表2
───────────────────────────────────
培養時間 16 18 20 40
───────────────────────────────────
喀痰検体 Viカンジダ 18(21) 23(24) 26(27) 26(27)
クロモカンジダ 2(21) 5(23) 15(25) 23(25)
膣検体 Viカンジダ 4(9) 14(16) 19(19) 19(19)
クロモカンジダ 0(8) 5(10) 5(19) 19(19)
───────────────────────────────────
実施例1で作成した培地を用いてカンジダ・アルビカンス、カンジダ・グラブラータ、カンジダ・トロピカリス、カンジダ・クルセイ、カンジダ・パラプシロシスを培養し、16,18,20,40時間後のコロニーの大きさや色について従来のクロモカンジダ培地を用いた場合と比較した。
Claims (4)
- 酵素基質として5−ブロム−4−クロロ−3−インドリルホスフェート.2Na(以下、x-phos.2Naという)および酸化還元試薬としてテトラゾリウムバイオレット(以下、TTVという)を含有することを特徴とするカンジダ鑑別用発色培地。
- x-phos.2Naの含有量が0.01〜0.1g/Lの範囲である請求項1記載のカンジダ鑑別用発色培地。
- TTVの含有量が0.001〜0.01g/Lの範囲である請求項1記載のカンジダ鑑別用発色培地。
- 培地1Lあたり、ペプトン10〜30g、酵母エキス1〜10g、白糖1〜20g、ブドウ糖1〜10g、TTV0.001〜0.01g、x-phos.2Na0.01〜0.1gおよび寒天3〜20gを含有する請求項1〜3のいずれか1項に記載のカンジダ鑑別用発色培地。
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JP2006025608A (ja) * | 2004-07-12 | 2006-02-02 | Chisso Corp | 微生物培地 |
JP6847628B2 (ja) * | 2015-10-29 | 2021-03-24 | 栄研化学株式会社 | カンジダ鑑別用発色培地 |
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