繰り返し凍結と解凍サイクルが,糞便の微生物群組成の安定性に及ぼす影響
Matteo Sangermani1,2, Indri Desiati3, Nicole Quattrini4
1HUNT Center for Molecular and Clinical Epidemiology, Department of Public Health and Nursing, Norwegian University of Science and Technology, Trondheim, Norway. matteo.sangermani@ntnu.no.
Scientific reports
|February 19, 2026
まとめ
保存された排泄物のサンプルを再分析することは,繰り返し凍結解凍サイクルが腸内微生物群の整合性に最小限の影響を与えるため,実行可能である. 個体間の差異は凍結解凍効果を上回り,微生物研究のためにサンプルの再利用を支えています.
科学分野:
- 微生物学 微生物学とは
- ゲノミクスゲノミクスとは
- バイオバンキング バイオバンキング
背景:
- 以前に採取したサンプルを再分析することで,新しいコホート研究と比較して時間と資源を節約できます.
- 繰り返し凍結解凍 (FT) サイクルを繰り返した後,糞便の微生物群の安定性について懸念があり,結果にバイアスを引き起こす可能性があります.
- FTサイクルが16S rRNA遺伝子配列に与える影響を調査することは,信頼性の高い微生物群分析に不可欠です.
研究 の 目的:
- 16S rRNA遺伝子配列解析で測定された糞便の微生物組成に繰り返し凍結解凍 (FT) サイクルが与える影響を評価する.
- FTサイクルが腸内微生物の分析に有意なバイアスを導入するかどうかを判断する.
- FTサイクルを施した保存された排泄物のサンプルの使用に関するガイドラインを提供すること.
主な方法:
- 16S rRNA遺伝子配列解析を用いて,最大6回のFTサイクルの排便サンプルへの影響を調査した.
- MaAsLin2やALDEx2.2など,差異的多量分析ツールを使用した.
- 新鮮で冷凍されていないサンプルと,繰り返しFTサイクルを受けたサンプルを比較した微生物の組成.
主要な成果:
- 個別サンプル間の変動は,FTサイクルの効果を一貫して上回った.
- MaAsLin2は,最初のFTサイクル後に限られた微生物の遺伝子の変化を特定し,サイクルの4〜6後にわずかな漸進的なシフトがあった.
- より保守的なALDEx2方法では,FTサイクル全体で有意な変化は検出されなかった.
- 最初の3つのFTサイクルは,全体的な微生物群組成に最小限の影響を及ぼしました.
結論:
- 1回の解凍を受けた保存された排泄物のサンプルの再利用は支持されます.
- 腸内微生物群の整合性を損なうリスクは,単一のFTサイクルを受けたサンプルでは最小限です.
- FTサイクルは,特に保守的な統計的手法では,微生物群分析に限られた影響を及ぼします.
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