A Multiplex Recombinase-Aided qPCR Assay for Highly Sensitive and Rapid Detection of khe, bla <subscript>KPC</subscript> <subscript>-2</subscript> , and bla <subscript>NDM</subscript> <subscript>-1</subscript> Genes in Klebsiella pneumoniae.

Hua, SW ; Wang, J ; et al.

In: Journal of clinical laboratory analysis, Jg. 38 (2024-05-01), Heft 9, S. e25038

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Objective: This study aimed to establish a highly sensitive and rapid single-tube, two-stage, multiplex recombinase-aided qPCR (mRAP) assay to specifically detect the khe, bla KPC-2 , and bla NDM-1 genes in Klebsiella pneumoniae.

Methods: mRAP was carried out in a qPCR instrument within 1 h. The analytical sensitivities of mRAP for khe, bla KPC-2 , and bla NDM-1 genes were tested using recombinant plasmids and dilutions of reference strains. A total of 137 clinical isolates and 86 sputum samples were used to validate the clinical performance of mRAP.

Results: mRAP achieved the sensitivities of 10, 8, and 14 copies/reaction for khe, bla KPC-2 , and bla NDM-1 genes, respectively, superior to qPCR. The Kappa value of qPCR and mRAP for detecting khe, bla KPC-2 , and bla NDM-1 genes was 1, 0.855, and 1, respectively (p < 0.05).

Conclusion: mRAP is a rapid and highly sensitive assay for potential clinical identification of khe, bla KPC-2 , and bla NDM-1 genes in K. pneumoniae.

Titel:	A Multiplex Recombinase-Aided qPCR Assay for Highly Sensitive and Rapid Detection of khe, bla <subscript>KPC</subscript> <subscript>-2</subscript> , and bla <subscript>NDM</subscript> <subscript>-1</subscript> Genes in Klebsiella pneumoniae.
Autor/in / Beteiligte Person:	Hua, SW ; Wang, J ; Zhao, ZJ ; Tian, FY ; Zhao, M ; Wang, YX ; Zhang, RQ ; Han, ZQ ; Gao, SJ ; Lv, XN ; Li, HY ; Shen, XX ; Ma, XJ ; Feng, ZS
Link:	Full Text Finder (Volltext)
Zeitschrift:	Journal of clinical laboratory analysis, Jg. 38 (2024-05-01), Heft 9, S. e25038
Veröffentlichung:	Hoboken, N.J. : Wiley ; <i>Original Publication</i>: New York : Alan R. Liss, Inc., c1987-, 2024
Medientyp:	academicJournal
ISSN:	1098-2825 (electronic)
DOI:	10.1002/jcla.25038
Schlagwort:	Humans Klebsiella Infections microbiology Klebsiella Infections diagnosis Sensitivity and Specificity Real-Time Polymerase Chain Reaction methods Bacterial Proteins genetics Recombinases genetics Recombinases metabolism Klebsiella pneumoniae genetics Klebsiella pneumoniae isolation & purification beta-Lactamases genetics Multiplex Polymerase Chain Reaction methods
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article Language: English [J Clin Lab Anal] 2024 May; Vol. 38 (9), pp. e25038. <i>Date of Electronic Publication: </i>2024 Apr 08. MeSH Terms: Klebsiella pneumoniae* / genetics ; Klebsiella pneumoniae* / isolation & purification ; beta-Lactamases* / genetics ; Multiplex Polymerase Chain Reaction* / methods ; Humans ; Klebsiella Infections / microbiology ; Klebsiella Infections / diagnosis ; Sensitivity and Specificity ; Real-Time Polymerase Chain Reaction / methods ; Bacterial Proteins / genetics ; Recombinases / genetics ; Recombinases / metabolism References: Front Microbiol. 2021 Jul 22;12:691289. (PMID: 34367092) ; J Sci Food Agric. 2022 Jul;102(9):3879-3886. (PMID: 34936095) ; J Clin Lab Anal. 2023 Mar;37(5):e24858. (PMID: 36916770) ; mSystems. 2022 Jun 28;7(3):e0129721. (PMID: 35574716) ; Emerg Infect Dis. 2020 Feb;26(2):289-297. (PMID: 31961299) ; Trop Med Infect Dis. 2023 Feb 06;8(2):. (PMID: 36828521) ; Front Cell Infect Microbiol. 2021 Sep 20;11:746325. (PMID: 34616692) ; Microbiol Mol Biol Rev. 2016 Jun 15;80(3):629-61. (PMID: 27307579) ; Lancet Infect Dis. 2017 Feb;17(2):153-163. (PMID: 27866944) ; Emerg Microbes Infect. 2022 Dec;11(1):1936-1949. (PMID: 35844192) ; J Infect Dis. 2017 Feb 15;215(suppl_1):S28-S36. (PMID: 28375512) ; Front Mol Biosci. 2022 Feb 09;8:794961. (PMID: 35223985) ; Front Cell Infect Microbiol. 2023 Jan 12;12:1000445. (PMID: 36710975) ; Front Bioeng Biotechnol. 2021 Nov 05;9:766411. (PMID: 34805120) ; Clin Infect Dis. 2018 Nov 13;67(suppl_2):S196-S205. (PMID: 30423057) ; Ann Clin Microbiol Antimicrob. 2013 Oct 22;12:30. (PMID: 24143953) ; EBioMedicine. 2017 May;19:98-106. (PMID: 28479289) ; Antimicrob Agents Chemother. 2020 Apr 21;64(5):. (PMID: 32094139) ; Front Cell Infect Microbiol. 2020 Jul 03;10:314. (PMID: 32719751) ; Int J Mol Sci. 2021 Apr 14;22(8):. (PMID: 33919847) ; Front Microbiol. 2023 Apr 27;14:1141424. (PMID: 37180280) ; EBioMedicine. 2020 Jan;51:102599. (PMID: 31911273) ; J Clin Microbiol. 2012 Aug;50(8):2596-600. (PMID: 22622443) ; Virulence. 2017 May 19;8(4):460-469. (PMID: 27593176) ; Biomed Res Int. 2022 Aug 8;2022:8612933. (PMID: 35978630) ; J Clin Diagn Res. 2014 Dec;8(12):DC23-6. (PMID: 25653949) ; Emerg Microbes Infect. 2020 Dec;9(1):1771-1779. (PMID: 32689907) ; Front Cell Infect Microbiol. 2022 May 19;12:877649. (PMID: 35663473) ; Emerg Infect Dis. 2010 Sep;16(9):1349-56. (PMID: 20735917) Grant Information: 2021YFC2301102 National Key R&D Program of China; 82202593 National Natural Science Foundation of China Contributed Indexing: Keywords: Carbapenem‐resistant Klebsiella pneumoniae (CRKP); RAA; blaKPC‐2; blaNDM‐1; khe Substance Nomenclature: EC 3.5.2.6 (beta-Lactamases) ; EC 3.5.2.6 (beta-lactamase NDM-1) ; 0 (Bacterial Proteins) ; 0 (Recombinases) Entry Date(s): Date Created: 20240409 Date Completed: 20240530 Latest Revision: 20240606 Update Code: 20240606 PubMed Central ID: PMC11137842

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