Rapid detection and identification of metallo-beta-lactamase-encoding genes by multiplex real-time PCR assay and melt curve analysis

Rapid detection and identification of metallo-beta-lactamase-encoding genes by multiplex real-time PCR assay and melt curve analysis

Author Mendes, Rodrigo E. Autor UNIFESP Google Scholar
Kiyota, Katia A. Autor UNIFESP Google Scholar
Monteiro, Jussimara Autor UNIFESP Google Scholar
Castanheira, Mariana Google Scholar
Andrade, Soraya S. Autor UNIFESP Google Scholar
Gales, Ana C. Autor UNIFESP Google Scholar
Pignatari, Antonio C. C. Google Scholar
Tufik, Sergio Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
AFIP
Abstract Metallo-p-lactamase enzymes (MOL) 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 MOL-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 (T.). the real-time PCR assay was able to detect all M beta L-harboring clinical isolates, and the T-m-assigned genotypes were 100% coincident with previous sequencing results. This assay could be suitable for identification of MOL-producing gram-negative bacteria by molecular diagnostic laboratories.
Language English
Date 2007-02-01
Published in Journal of Clinical Microbiology. Washington: Amer Soc Microbiology, v. 45, n. 2, p. 544-547, 2007.
ISSN 0095-1137 (Sherpa/Romeo, impact factor)
Publisher Amer Soc Microbiology
Extent 544-547
Origin http://dx.doi.org/10.1128/JCM.01728-06
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000244270000045
URI http://repositorio.unifesp.br/handle/11600/29478

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