Multiplexed detection of bacterial nucleic acids using Cas13 in droplet microarrays.

التفاصيل البيبلوغرافية
العنوان:	Multiplexed detection of bacterial nucleic acids using Cas13 in droplet microarrays.
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.)
References:	ACS Infect Dis. 2018 Sep 14;4(9):1278-1282. (PMID: 30113801) Nucleic Acids Res. 2019 Aug 22;47(14):e83. (PMID: 31114866) J Clin Microbiol. 2013 May;51(5):1528-33. (PMID: 23486707) Nat Microbiol. 2019 Apr;4(4):663-674. (PMID: 30742071) Nat Med. 2019 Dec;25(12):1858-1864. (PMID: 31768064) Appl Environ Microbiol. 2014 Aug;80(16):5116-23. (PMID: 24928874) Nat Rev Microbiol. 2017 Oct 12;15(11):697-703. (PMID: 29021600) Nat Commun. 2018 Aug 21;9(1):3347. (PMID: 30131493) J Microbiol Methods. 2007 May;69(2):330-9. (PMID: 17391789) BMC Bioinformatics. 2004 Aug 19;5:113. (PMID: 15318951) Science. 2017 Apr 28;356(6336):438-442. (PMID: 28408723) Sci Transl Med. 2020 Jun 3;12(546):. (PMID: 32493791) Front Microbiol. 2012 Jan 23;3:17. (PMID: 22319519) Proc Natl Acad Sci U S A. 2018 Jun 26;115(26):6685-6690. (PMID: 29899149) Science. 2018 Apr 27;360(6387):436-439. (PMID: 29449511) Euro Surveill. 2020 Jan;25(3):. (PMID: 31992387) Nat Med. 2020 Mar;26(3):333-340. (PMID: 32066974) Nature. 2020 Jun;582(7811):277-282. (PMID: 32349121) Front Microbiol. 2018 May 29;9:1097. (PMID: 29896172) Science. 2018 Apr 27;360(6387):439-444. (PMID: 29449508) Proc Natl Acad Sci U S A. 2019 Jun 25;116(26):12804-12809. (PMID: 31186361) Nucleic Acids Res. 2014 Jan;42(Database issue):D625-32. (PMID: 24198250) BMC Bioinformatics. 2018 Sep 29;19(1):343. (PMID: 30268091) N Engl J Med. 2020 Aug 20;383(8):e56. (PMID: 32767891) Anal Chem. 2013 Aug 20;85(16):7629-36. (PMID: 23763357) Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14676-9. (PMID: 8962113) Sci Rep. 2019 Mar 14;9(1):4516. (PMID: 30872641) Nat Biotechnol. 2022 Jul;40(7):1123-1131. (PMID: 35241837) Lancet Respir Med. 2016 Mar;4(3):213-24. (PMID: 26907218) Nat Biotechnol. 2017 Jul;35(7):676-683. (PMID: 28604660) N Engl J Med. 2012 Feb 2;366(5):454-61. (PMID: 22296079) Science. 2018 Apr 27;360(6387):444-448. (PMID: 29700266)
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

Full Text Finder

الوصف
تدمد:	2752-6542
DOI:	10.1093/pnasnexus/pgac021