使用CRISPR-Cas13进行现场部署的病毒诊断
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
概括
一个新的诊断平台,SHERLOCK (特定高灵敏度酶记者解锁),可以快速检测患者样本中的寨卡病毒 (ZIKV) 和登革热病毒 (DENV). 通过HUDSON (加热未提取的诊断样本以消灭核酶) 协议,可以在不到两个小时的时间内进行无仪器检测.
科学领域:
- 分子生物学
- 病毒学
- 生物技术
背景情况:
- 全球传染病缓解需要敏感,具体和现场部署的诊断工具.
- 现有的诊断方法可能缺乏迅速应对疫情所需的速度和可访问性.
研究的目的:
- 评估基于Cas13的SHERLOCK平台对寨卡病毒 (ZIKV) 和登革热病毒 (DENV) 的敏感和特异性检测.
- 开发和验证HUDSON协议,直接从患者样本中快速检测病毒.
- 评估SHERLOCK在区分DENV血清型和ZIKV菌株方面的能力.
主要方法:
- 使用基于Cas13的Sherlock平台进行核酸检测.
- 开发并实施HUDSON准备样本的协议.
- 对ZIKV和DENV的测试灵敏度降至每微升1副本.
- 证实了谢洛克能够区分DENV血清型和ZIKV菌株.
主要成果:
- SHERLOCK检测到ZIKV和DENV的度只有1副本/μL.
- 通过HUDSON协议,可以在不到2小时的时间内从患者样本中检测DENV.
- 谢洛克成功区分了四种DENV血清和流行性ZIKV菌株.
- 实现了快速 (< 1 周) 设计和测试病毒单核酸多态性的无仪器测试.
结论:
- SHERLOCK平台与HUDSON协议相结合,为ZIKV和DENV提供了灵敏,具体和快速的诊断解决方案.
- 这种方法有助于无仪器检测,这对于管理传染病爆发至关重要.
- 该平台能够检测特定的病毒变异和单核酸多态性,从而提高其在分子流行病学中的实用性.
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