JP6769872B2 - 代謝測定によるマススペクトル耐性判定 - Google Patents
代謝測定によるマススペクトル耐性判定 Download PDFInfo
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Description
更に次に、この阻害剤を阻害する物質を用いて、過剰な阻害剤を中和することも可能である。
抗生物質を含まない培地での培養(108)、ならびに、抗生物質Ab1 (109) およびAb2 (110) を含んだ2つの培養である。この例において、培地にはすでにインジケーターが含まれている。もちろん、更に他の抗生物質を入れた培養物も調製することができ、また耐性の強度も判定する場合には、異なる抗生物質濃度での培養物も調製することができる。微生物に衝撃を与えないよう、また加熱によって時間の遅れが生じないよう、培養物はすべて、最適温度で調製済みである。培養時間は、待ち時間および微生物の倍加時間(世代期間) に依存し、これは微生物の同定により既知である。培養は、倍加時間1回〜3回分だけが必要である。成長の速い微生物には、約20〜40分間で十分である。
Claims (20)
- 微生物の抗生物質に対する耐性をマススペクトルにより判定する方法であって、
前記微生物を前記抗生物質とともに予備培養した後、
少なくとも1種類の栄養成分を新たに添加した、特定の濃度の前記抗生物質を含む合成培地で前記微生物を培養し、
前記合成培地中の前記添加された栄養成分が減少したかどうか、または、前記添加された栄養成分の化学的変性変異体が増加したかどうかを、マススペクトルにより測定し、
この結果、前記添加された栄養成分が減少した場合、あるいは前記添加された栄養成分の化学的変性変異体が増加した場合に、前記特定の濃度の抗生物質対して前記微生物が耐性であると判定する、方法。 - 前記添加された栄養成分の減少、又は前記添加された栄養成分の化学的変性変異体の増加が、所定量添加された参照物質と比較することにより判定される、請求項1に記載の方法。
- 前記参照物質が、D-アミノ酸から作られたペプチドである、請求項2に記載の方法。
- 前記参照物質が、前記合成培地中での成長の完了後に所定量添加される、請求項2に記載の方法。
- 前記添加された栄養成分がペプチドであり、前記栄養成分の添加の直後に、既に分泌されているペプチダーゼの阻害剤が添加される、請求項1に記載の方法。
- 耐性の強度を判定するため、前記微生物を、いくつかの濃度の前記抗生物質を含んだ合成培地で培養する、請求項1に記載の方法。
- 微生物の抗生物質に対する耐性をマススペクトルにより判定する方法であって、
前記微生物を、特定の濃度の抗生物質と、D-アミノ酸のコアを含む少なくとも1種類のペプチドとを含む培地で培養し、
前記D-アミノ酸のコアのみを含むペプチドが増加するかどうかを、マススペクトルで測定し、
前記D-アミノ酸のコアのみを含むペプチドの増加があった場合に、前記特定の濃度の抗生物質対して前記微生物が耐性であると判定する、方法。 - 前記少なくとも1種類のペプチドが同位体標識されたアミノ酸からなり、培地中で長さが短くなった同位体標識ペプチドの生成が、マススペクトルで測定される、請求項7に記載の方法。
- 前記増加は、所定量添加された少なくとも1種類の参照物質と比較することにより判定される、請求項7に記載の方法。
- 前記参照物質が、D-アミノ酸から作られたペプチドである、請求項9に記載の方法。
- 前記参照物質が、前記培地中での成長の完了後に所定量添加される、請求項9に記載の方法。
- 前記微生物が、抗生物質を含む第1の合成培地で予備培養されてから、この予備培養された微生物が更に第2の合成培地で培養される、請求項7に記載の方法。
- 前記微生物が、抗生物質で予備培養されてから、前記少なくとも1種類のペプチドが添加される、請求項7に記載の方法。
- 前記微生物を分割して、これらの分割部分を、抗生物質を含まない第1の合成培地と、抗生物質を含む第2の合成培地とで、それぞれ培養する、請求項7に記載の方法。
- マトリックス支援レーザー脱離/イオン化(MALDI)またはエレクトロ・スプレーイオン化 (ESI)のいずれかの方法が、マススペクトル測定におけるイオン化方法として用いられる、請求項1または請求項7に記載の方法。
- 前記合成培地は、前記添加された少なくとも1種類の栄養成分として、少なくとも1種類のペプチドを含む、請求項1に記載の方法。
- 前記合成培地は、個別のアミノ酸を含まない、請求項16に記載の方法。
- 前記少なくとも1種類のペプチドは、全20種類のアミノ酸を含む、請求項16に記載の方法。
- 前記抗生物質の濃度は、抗生物質の最小阻害濃度(MIC)の10倍である、請求項1または請求項7に記載の方法。
- 微生物の抗生物質に対する耐性をマススペクトルにより判定する方法であって、
前記微生物を、特定の濃度の抗生物質を含む第1の合成培地で予備培養し、
前記予備培養された微生物を、特定の濃度の前記抗生物質を含む第2の合成培地で更に培養し、
前記第2の合成培地の栄養成分の分解、または、前記第2の合成培地の栄養成分の化学的変性変異体の増加を、マススペクトルで測定し、
この結果、前記栄養成分が減少した場合、あるいは前記栄養成分の化学的変性変異体が増加した場合、前記特定の濃度の前記抗生物質に対して前記微生物が耐性であると判定する、方法。
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PCT/EP2015/050526 WO2015107054A1 (en) | 2014-01-17 | 2015-01-14 | Mass spectrometric resistance determination by measuring metabolism |
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WO2019108125A1 (en) | 2017-11-30 | 2019-06-06 | Ascilion Ab | A method for determining microbial susceptibility to antibiotic agents |
EP3717657A1 (en) | 2017-11-30 | 2020-10-07 | Ascilion AB | A system for assessing microbial susceptibility to antibiotic agents |
US20220235392A1 (en) * | 2019-05-31 | 2022-07-28 | Lewis Ian Andrew | Metabolomic Characterization of Microorganisms |
CN112051321B (zh) * | 2020-08-24 | 2022-02-15 | 复旦大学 | 结合氘水培养和基质辅助激光解吸电离飞行时间质谱分析的快速抗生素敏感性测试方法 |
KR20230045361A (ko) * | 2021-09-28 | 2023-04-04 | 주식회사 엘지화학 | 미생물 배양 농도 예측 방법 및 시스템 |
DE102021130356B3 (de) | 2021-11-19 | 2023-04-20 | Bruker Daltonics GmbH & Co. KG | Referenzdatensatz-basiertes, spektrometrisches Charakterisieren von Zellsubstraten unter Verwendung von Teilbibliotheken |
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FR2671100B1 (fr) * | 1990-12-28 | 1993-03-05 | Bio Merieux | Procede d'analyse bacteriologique, et milieu de detection des bacteries genre salmonella. |
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