دورية أكاديمية
Multiplexed detection of bacterial nucleic acids using Cas13 in droplet microarrays.
العنوان: | Multiplexed detection of bacterial nucleic acids using Cas13 in droplet microarrays. |
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المؤلفون: | Thakku SG; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Ackerman CM; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Myhrvold C; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Bhattacharyya RP; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Livny J; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Ma P; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Gomez GI; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Sabeti PC; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Blainey PC; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Hung DT; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. |
المصدر: | PNAS nexus [PNAS Nexus] 2022 Apr 15; Vol. 1 (1), pp. pgac021. Date of Electronic Publication: 2022 Apr 15 (Print Publication: 2022). |
نوع المنشور: | Journal Article |
اللغة: | English |
بيانات الدورية: | Publisher: Oxford University Press on behalf of the National Academy of Sciences Country of Publication: England NLM ID: 9918367777906676 Publication Model: eCollection Cited Medium: Internet ISSN: 2752-6542 (Electronic) Linking ISSN: 27526542 NLM ISO Abbreviation: PNAS Nexus Subsets: PubMed not MEDLINE |
أسماء مطبوعة: | Original Publication: [Oxford] : Oxford University Press on behalf of the National Academy of Sciences, [2022]- |
مستخلص: | 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. (© The Author(s) 2022. Published by Oxford University Press on behalf of the National Academy of Sciences.) |
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معلومات مُعتمدة: | U19 AI142780 United States AI NIAID NIH HHS |
تواريخ الأحداث: | Date Created: 20220422 Latest Revision: 20220716 |
رمز التحديث: | 20221213 |
مُعرف محوري في PubMed: | PMC9013781 |
DOI: | 10.1093/pnasnexus/pgac021 |
PMID: | 35450424 |
قاعدة البيانات: | MEDLINE |
تدمد: | 2752-6542 |
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DOI: | 10.1093/pnasnexus/pgac021 |