JP2020517300A - 抗生物質感受性の全同定のためのアッセイ - Google Patents
抗生物質感受性の全同定のためのアッセイ Download PDFInfo
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- JP2020517300A JP2020517300A JP2020507493A JP2020507493A JP2020517300A JP 2020517300 A JP2020517300 A JP 2020517300A JP 2020507493 A JP2020507493 A JP 2020507493A JP 2020507493 A JP2020507493 A JP 2020507493A JP 2020517300 A JP2020517300 A JP 2020517300A
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- antibiotics
- antibiotic
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- bacteria
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Abstract
Description
本出願は、2017年4月19日に出願された米国仮出願第62/487,395号の利益を主張し、その内容全体は参照により本明細書に組み込まれる。
感染性疾患は複数の臓器系に影響を及ぼし、重大な罹患率、死亡率、および経済的影響の原因となっている。感染病原体は、単一の病原体としてではなく、複雑な複数菌感染症として最もよく存在している。体内では、これらの複数菌感染症は、生物が感受性である抗生物質の種類だけでなく、感染症の治療に必要な抗生物質のレベルおよび個々の病原体の毒性を変化させる相利共生によって互いに協働する。現在の最も基準となる検査は培養と感受性であり、サンプルを、寒天培地を含むプレート上に設置して一晩培養するプロセスは、個々の生物を分離および同定することを可能にする。通常、抗生物質感受性試験を通じて個々の生物がどの抗生物質に感受性であるかを同定するために、追加の日が必要である。このプロセスの有効性は、手法の感受性と特異性が本質的に低いために制限される。例えば、最近の研究では、すべての尿培養結果の最大25%が偽陰性所見を生み出し、すべての尿路病原体の最大2/3が培養によって失われていることがわかっている。このプロセスは、通常48〜72時間かかり、正確な診断なしに、および感染症の治療に有効な抗生物質の種類に関する適切な誘導なしに、臨床医が経験的に患者を治療することを余儀なくされるという点で、臨床診療に大きな影響を及ぼす。
バンコマイシン、TMP/SMX、トリメトプリム、スルファメトキサゾール、ニトロフラントイン、シプロフロキサシン、ガチフロキサシン、レボフロキサシン、モキシフロキサシン、オフロキサシン、プロリフロキサシン、クロリマイシン、アジスロマイシン、およびクラリスロマイシンの1つ以上である。
このアッセイでの処理に適した尿サンプルを、BDバキュテイナ(グレートップ)チューブまたは他の適切な漏れ防止滅菌容器を使用して、採取、輸送、保管する。尿サンプルを、試験結果が損なわれる前に、室温で48時間保持する場合がある。
例2.増殖培地を含む寒天を利用したABRアッセイの検証
例3.液体増殖培地を利用した抗生物質耐性(ABR)アッセイ
例4.液体増殖培地を利用したABRアッセイの検証
Claims (38)
- サンプル中の細菌の混合集団の複数の抗生物質に対する感受性を同時に決定するための方法であって、
前記サンプルのある量を複数の抗生物質含有培地に同時に適用し、前記サンプルが感染症が疑われる被験者からのものである工程と、
前記サンプルを前記培地上で一定期間培養する工程と、
前記培地に存在する細菌を溶液で懸濁し、前記懸濁液を回収する工程と、
前記懸濁液のOD600を測定し、懸濁液の調整されたOD600が前記サンプル中の細菌の個々の抗生物質に対する感受性または耐性を示す工程と、
前記感受性情報を医療専門家に提供して、前記被験者の抗生物質療法を決定する工程とを含む、方法。 - 前記サンプルが複数種の細菌を含む、請求項1に記載の方法。
- 前記複数の抗生物質含有培地が、複数のウェルに抗生物質含有細菌増殖培地が配置されたマルチウェルアッセイプレートを含む、請求項1に記載の方法。
- 前記マルチウェルアッセイプレートが96個のウェルを有する、請求項3に記載の方法。
- 前記細菌増殖培地が寒天含有培地である、請求項3に記載の方法。
- 前記複数の抗生物質が、スルファメトキサゾール、トリメトプリム、ニトロフラントイン、ホスホマイシン、アモキシシリン、クラブラナート、セフポドキシム、セフジニル、セファクロル、セフェピム、セファゾリン、シプロフロキサシン、レボフロキサシン、セフトリアキソン、ゲンタマイシン、トブラマイシン、アビバクタム、セフタジジム、セフトロザン、タゾバクタム、ノルフロキサシン、メロペネム、ピペラシリン、セフォキシチン、テトラサイクリン、スルバクタム、セフロキシム、およびバンコマイシンの2以上を含む、請求項1に記載の方法。
- 前記複数のウェルのそれぞれが、単一の抗生物質または抗生物質の組み合わせを含む、請求項6に記載の方法。
- 前記抗生物質の組み合わせが2つの抗生物質を含む、請求項7に記載の方法。
- 前記複数の抗生物質が、少なくとも3つの抗生物質、少なくとも4つの抗生物質、少なくとも5つの抗生物質、少なくとも6つの抗生物質、少なくとも7つの抗生物質、少なくとも8つの抗生物質、少なくとも9つの抗生物質、または少なくとも10の抗生物質を含む、請求項7に記載の方法。
- 各抗生物質または抗生物質の組み合わせが、2つ以上の濃度で前記ウェルに存在する、請求項1に記載の方法。
- 各抗生物質、または抗生物質の組み合わせが、3つ以上の濃度で前記ウェルに存在する、請求項10に記載の方法。
- 前記懸濁液のOD600が複数回測定される、請求項1に記載の方法。
- 前記OD600が2回、3回、4回、または5回測定される、請求項12に記載の方法。
- 前記調整されたOD600が、バックグラウンド対照を差し引いた複数の測定値の平均OD600を含む、請求項1に記載の方法。
- 前記調整されたOD600が0.025未満である場合、前記サンプル中の前記細菌が抗生物質または抗生物質の組み合わせに対して感受性である、請求項1に記載の方法。
- 前記調整されたOD600が0.025以上である場合、前記サンプル中の前記細菌が抗生物質または抗生物質の組み合わせに対して耐性である、請求項1に記載の方法。
- 前記サンプルが生物学的液体または生検サンプルである、請求項1に記載の方法。
- 前記サンプルが、尿、血液、唾液、喀痰、肺洗浄液、膣分泌物、生検組織、または脳脊髄液を含む、請求項17に記載の方法。
- サンプル中の細菌の混合集団の複数の抗生物質に対する感受性を同時に決定するための方法であって、
サンプルのある量を増殖培地と混合し、一定期間培養する工程と、
前記サンプルを複数の抗生物質含有培地に同時に適用し、前記サンプルが感染症が疑われる被験者からのものである工程と、
前記サンプルを前記抗生物質含有培地で一定期間培養する工程と、
前記培地中の細菌増殖をOD600測定により測定し、前記サンプルの調整されたOD600測定値が、前記サンプル内の前記細菌の個々の抗生物質に対する感受性または耐性を示す工程と、
前記感受性情報を医療専門家に提供して、前記被験者の抗生物質療法を決定する工程とを含む、方法。 - 前記サンプルが複数の種の細菌を含む、請求項19に記載の方法。
- 前記複数の抗生物質含有培地が、複数のウェルに抗生物質含有細菌増殖培地が配置されたマルチウェルアッセイプレートを含む、請求項19に記載の方法。
- 前記マルチウェルアッセイプレートが96個のウェルを有する、請求項19に記載の方法。
- 前記培地が液体ブロス培地である、請求項19に記載の方法。
- 前記培地がミューラーヒントン培地である、請求項23に記載の方法。
- 前記複数の抗生物質が、スルファメトキサゾール、トリメトプリム、ニトロフラントイン、ホスホマイシン、アモキシシリン、クラブラナート、セフポドキシム、セフジニル、セファクロル、シプロフロキサシン、レボフロキサシン、セフトリアキソン、ゲンタマイシン、トブラマイシン、アビバクタム、セフタジジム、セフトロザン、タゾバクタム、ノルフロキサシン、メロペネム、ピペラシリン、セフォキシチン、テトラサイクリン、スルバクタム、セフロキシム、およびバンコマイシンの2以上を含む、請求項19に記載の方法。
- 前記複数のウェルのそれぞれが、単一の抗生物質または抗生物質の組み合わせを含む、請求項26に記載の方法。
- 前記抗生物質の組み合わせが2つの抗生物質を含む、請求項26に記載の方法。
- 前記複数の抗生物質が、少なくとも3つの抗生物質、少なくとも4つの抗生物質、少なくとも5つの抗生物質、少なくとも6つの抗生物質、少なくとも7つの抗生物質、少なくとも8つの抗生物質、少なくとも9つの抗生物質、または少なくとも10の抗生物質を含む、請求項26に記載の方法。
- 各抗生物質、または抗生物質の組み合わせが、2つ以上の濃度で前記ウェルに存在する、請求項19に記載の方法。
- 各抗生物質、または抗生物質の組み合わせが、3つ以上の濃度で含まれる、請求項29に記載の方法。
- 前記サンプルのOD600が1回測定される、請求項19に記載の方法。
- 前記サンプルのOD600が複数回測定される、請求項19に記載の方法。
- 前記OD600が2回、3回、4回、または5回測定される、請求項34に記載の方法。
- 前記調整されたOD600が、バックグラウンド対照を差し引いた、単一のOD600測定値、または複数の測定値の平均OD600を含む、請求項19に記載の方法。
- 前記調整されたOD600が0.065未満である場合、前記サンプル中の前記細菌が抗生物質または抗生物質の組み合わせに対して感受性である、請求項19に記載の方法。
- 前記調整されたOD600が0.065以上である場合、前記サンプル中の細菌が抗生物質または抗生物質の組み合わせに対して耐性である、請求項19に記載の方法。
- 前記サンプルが生物学的液体または生検サンプルである、請求項19に記載の方法。
- 前記サンプルが、尿、血液、唾液、喀痰、肺洗浄液、膣分泌物、生検組織、または脳脊髄液を含む、請求項37に記載の方法。
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