CRISPR-Cas13を用いたフィールドデプロイ可能なウイルス診断
Cameron Myhrvold1,2, Catherine A Freije1,2,3, Jonathan S Gootenberg4,5,6,7,8
1Broad Institute of the Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA. pardis@broadinstitute.org cmyhrvol@broadinstitute.org cfreije@broadinstitute.org.
Science (New York, N.Y.)
|April 28, 2018
まとめ
新しい診断プラットフォームであるSHERLOCK (特殊高感度酵素レポーター解鎖) は,患者サンプルでジカウイルス (ZIKV) とデング熱ウイルス (DENV) を迅速に検出します. HUDSON (核酸を消去するために未抽出の診断サンプルを加熱する) プロトコルにより,計測器なしで2時間以内に検出できます.
科学分野:
- 分子生物学
- ウイルス学
- バイオテクノロジー
背景:
- 世界的な感染症の緩和には 敏感で特異的な フィールドデプロイ可能な診断ツールが必要です
- 現存する診断方法は,迅速なアウトブレイク対応に必要なスピードとアクセシビリティを欠いている可能性があります.
研究 の 目的:
- ジカウイルス (ZIKV) とデング熱ウイルス (DENV) の敏感で特異的な検出のためのCas13ベースのSHERLOCKプラットフォームを評価する.
- 患者のサンプルから直接,素早く,器具なしでウイルスを検出するためのハドソンプロトコルを開発し,検証する.
- SHERLOCKのDENV血清型とZIKV株の区別能力を評価する.
主な方法:
- 核酸検出のCas13ベースのSherlockプラットフォームを使用しました.
- 試料の準備のためのハドソンプロトコルを開発し,実装しました.
- ZIKV と DENV に対して,微リットルあたり1コピーの検査感度を下げました.
- DENVの血清型とZIKVの株を区別する能力が検証された.
主要な成果:
- SHERLOCKは,ZIKVとDENVを1コピー/μLの濃度で検出しました.
- HUDSONプロトコルにより,患者のサンプルから 2時間未満で DENVの検出が可能になりました.
- SHERLOCKは4つのDENV血清型とパンデミックZIKV株を成功裏に区別しました.
- ウイルス単核型多形性に対する無器具検査の迅速な (< 1 週間) 設計と試験が達成されました.
結論:
- SHERLOCKプラットフォームは,HUDSONプロトコルと組み合わせて,ZIKVとDENVの敏感で,特定の,迅速な診断ソリューションを提供します.
- このアプローチは 感染症の発生管理に不可欠な 医療現場での検査を 容易にします
- このプラットフォームは,特定のウイルス変種と単一核性多型性を検出する能力があり,分子疫学における有用性を高めています.
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