Rapid Detection and Identification of Metallo-β-Lactamase-Encoding Genes by Multiplex Real-Time PCR Assay and Melt Curve Analysis
Rodrigo E. Mendes,Rodrigo E. Mendes,Katia A. Kiyota,Jussimara Monteiro,Jussimara Monteiro,Mariana Castanheira,Soraya Sgambatti Andrade,Soraya Sgambatti Andrade,Ana Cristina Gales,Antonio Carlos Campos Pignatari,Sergio Tufik,Sergio Tufik +11 more
TLDR
The real-time PCR assay was able to detect all MβL-harboring clinical isolates, and the Tm-assigned genotypes were 100% coincident with previous sequencing results, meaning this assay could be suitable for identification of M βL-producing gram-negative bacteria by molecular diagnostic laboratories.Abstract:
Metallo-β-lactamase enzymes (MβL) are encoded by transferable genes, which appear to spread rapidly among gram-negative bacteria The objective of this study was to develop a multiplex real-time PCR assay followed by a melt curve step for rapid detection and identification of genes encoding MβL-type enzymes based on the amplicon melting peak The reference sequences of all genes encoding IMP and VIM types, SPM-1, GIM-1, and SIM-1 were downloaded from GenBank, and primers were designed to obtain amplicons showing different sizes and melting peak temperatures (Tm) The real-time PCR assay was able to detect all MβL-harboring clinical isolates, and the Tm-assigned genotypes were 100% coincident with previous sequencing results This assay could be suitable for identification of MβL-producing gram-negative bacteria by molecular diagnostic laboratoriesread more
Citations
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Alexandre R. Marra,Alexandre R. Marra,Luis Fernando Aranha Camargo,Luis Fernando Aranha Camargo,Antonio Carlos Campos Pignatari,Teresa Sukiennik,Paulo Renato Petersen Behar,Eduardo Alexandrino Servolo Medeiros,Julival Ribeiro,Evelyne Santana Girão,Luci Correa,Carla Morales Guerra,Carlos Brites,Carlos Alberto Pires Pereira,Irna Carla do Rosário Souza Carneiro,Marise Reis,Marta Antunes de Souza,Regina Tranchesi,Cristina U. Barata,Michael B. Edmond +19 more
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References
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Metallo-β-Lactamases: the Quiet before the Storm?
TL;DR: Their rapid dissemination is worrisome and necessitates the implementation of not just surveillance studies but also metallo-β-lactamase inhibitor studies securing the longevity of important anti-infectives.
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Molecular characterization of an enterobacterial metallo beta-lactamase found in a clinical isolate of Serratia marcescens that shows imipenem resistance.
E Osano,Yoshichika Arakawa,R Wacharotayankun,Michio Ohta,Toshinobu Horii,Hideo Ito,F Yoshimura,Nobuo Kato +7 more
TL;DR: Results clearly show that IMP-1 is an enterobacterial metallo beta-lactamase, of which the primary structure has been completely determined, that confers resistance to carbapenems and other broad-spectrum beta- lactams.
Journal ArticleDOI
Novel Acquired Metallo-β-Lactamase Gene, blaSIM-1, in a Class 1 Integron from Acinetobacter baumannii Clinical Isolates from Korea
Kyungwon Lee,Jong Hwa Yum,Dongeun Yong,Hyuk Lee,Heung Dong Kim,Jean Denis Docquier,Gian Maria Rossolini,Yunsop Chong +7 more
TL;DR: The blaSIM-1 gene was carried on a gene cassette inserted into a class 1 integron, which included three additional cassettes (arr-3, catB3, and aadA1).
Journal ArticleDOI
Molecular characterization of SPM-1, a novel metallo-β-lactamase isolated in Latin America: report from the SENTRY antimicrobial surveillance programme
Mark Toleman,Alan M. Simm,Tanya A. Murphy,Ana Cristina Gales,Doug Biedenbach,Ronald N. Jones,Timothy R. Walsh +6 more
TL;DR: SPM-1 is a distinctly different metallo-beta-lactamase from VIM and IMP and, accordingly, represents a new subfamily of mobile metallo,beta, and lactamases, and possesses a unique loop of 23 residues that accounts for the higher molecular mass.
Journal ArticleDOI
Molecular characterization of a beta-lactamase gene, blaGIM-1, encoding a new subclass of metallo-beta-lactamase.
TL;DR: Kinetics analysis revealed that GIM-1 has no clear preference for any substrate and did not hydrolyze azlocillin, aztreonam, and the serine-β-lactamase inhibitors, and represents the fourth subclass of mobile MβL enzymes to be characterized.