Multiplexed detection of bacterial nucleic acids using Cas13 in droplet microarrays.
Multiplexed detection of bacterial nucleic acids using Cas13 in droplet microarrays.
复制标题
在液滴微阵列中使用Cas13的细菌核酸的多重检测。
DOI：
10.1093/pnasnexus/pgac021
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发表时间：
2022-03
期刊：
影响因子：
--
通讯作者：
Hung, Deborah T.
中科院分区：
文献类型：
--
作者：
Thakku, Sri Gowtham;Ackerman, Cheri M.;Myhrvold, Cameron;Bhattacharyya, Roby P.;Livny, Jonathan;Ma, Peijun;Gomez, Giselle Isabella;Sabeti, Pardis C.;Blainey, Paul C.;Hung, Deborah T.
快速和准确的感染诊断是个体患者护理和公共卫生管理的基础。核酸检测方法对这一努力至关重要，但在目标病原体的广度或所需的专业知识和基础设施方面受到限制。我们在这里提出了一种能够快速鉴定细菌病原体的高通量系统bCARMEN，其利用：（1）具有增强的灵敏度和特异性的基于CRISPR-Cas 13的模块化核酸检测;和（2）液滴微流体系统，其能够以纳升体积进行数千次同时的空间多路检测反应;和（3）进一步提高灵敏度和特异性的新的预扩增策略。我们证明bCARMEN能够检测和区分52种临床相关的细菌物种和几个关键的抗生素耐药基因。我们进一步开发了一种简单的原理工作流程证明，使用稳定的试剂和手机摄像头光学读数，开辟了快速护理点多重细菌病原体鉴定和抗生素敏感性测试的可能性。
Rapid and accurate diagnosis of infections is fundamental to individual patient care and public health management. Nucleic acid detection methods are critical to this effort, but are limited either in the breadth of pathogens targeted or by the expertise and infrastructure required. We present here a high-throughput system that enables rapid identification of bacterial pathogens, bCARMEN, which utilizes: (1) modular CRISPR-Cas13-based nucleic acid detection with enhanced sensitivity and specificity; and (2) a droplet microfluidic system that enables thousands of simultaneous, spatially multiplexed detection reactions at nanoliter volumes; and (3) a novel preamplification strategy that further enhances sensitivity and specificity. We demonstrate bCARMEN is capable of detecting and discriminating 52 clinically relevant bacterial species and several key antibiotic resistance genes. We further develop a simple proof of principle workflow using stabilized reagents and cell phone camera optical readout, opening up the possibility of a rapid point-of-care multiplexed bacterial pathogen identification and antibiotic susceptibility testing.
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DOI：
10.1126/science.aas8836
发表时间：
2018-04-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Myhrvold C;Freije CA;Gootenberg JS;Abudayyeh OO;Metsky HC;Durbin AF;Kellner MJ;Tan AL;Paul LM;Parham LA;Garcia KF;Barnes KG;Chak B;Mondini A;Nogueira ML;Isern S;Michael SF;Lorenzana I;Yozwiak NL;MacInnis BL;Bosch I;Gehrke L;Zhang F;Sabeti PC
通讯作者：
Sabeti PC
影响因子：
64.8
作者：
Ackerman, Cheri M.;Myhrvold, Cameron;Sabeti, Pardis C.
通讯作者：
Sabeti, Pardis C.
影响因子：
46.9
作者：
Metsky, Hayden C.;Welch, Nicole L.;Pillai, Priya P.;Haradhvala, Nicholas J.;Rumker, Laurie;Mantena, Sreekar;Zhang, Yibin B.;Yang, David K.;Ackerman, Cheri M.;Weller, Juliane;Blainey, Paul C.;Myhrvold, Cameron;Mitzenmacher, Michael;Sabeti, Pardis C.
通讯作者：
Sabeti, Pardis C.
影响因子：
4.6
作者：
Bhattacharyya, Roby P.;Walker, Mark;Hung, Deborah T.
通讯作者：
Hung, Deborah T.
影响因子：
5.2
作者：
Martino, Amanda J.;Rhodes, Matthew E.;House, Christopher H.
通讯作者：
House, Christopher H.