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Primer Evaluation for PCR and its Application for Detection of Carbapenemases in Enterobacteriaceae
P. Mlynarcik, M. Roderova, M. Kolar,
Jazyk angličtina Země Írán
Typ dokumentu časopisecké články
NLK
Free Medical Journals
od 2008
PubMed Central
od 2014 do 2016
Europe PubMed Central
od 2014 do 2016
ProQuest Central
od 2008-10-01
Open Access Digital Library
od 2014-01-01
Health & Medicine (ProQuest)
od 2008-10-01
ROAD: Directory of Open Access Scholarly Resources
od 2008
PubMed
27099689
DOI
10.5812/jjm.29314
Knihovny.cz E-zdroje
- Publikační typ
- časopisecké články MeSH
BACKGROUND: During the last decade, the prevalence of carbapenem-resistant Enterobacteriaceae in human patients has increased. Carbapenemase-producing bacteria are usually multidrug resistant. Therefore, early recognition of carbapenemase producers is critical to prevent their spread. OBJECTIVES: The objective of this study was to develop the primers for single and/or multiplex PCR amplification assays for simultaneous identification of class A, class B, and class D carbapenem hydrolyzing β-lactamases in Enterobacteriaceae and then to evaluate their efficiency. MATERIALS AND METHODS: The reference sequences of all genes encoding carbapenemases were downloaded from GenBank. Primers were designed to amplify the following 11 genes: bla KPC, bla OXA, bla VIM, bla NDM, bla IMP, bla SME, bla IMI, bla GES , bla GIM, bla DIM and bla CMY . PCR conditions were tested to amplify fragments of different sizes. Two multiplex PCR sets were created for the detection of clinically important carbapenemases. The third set of primers was included for detection of all known carbapenemases in Enterobacteriaceae. They were evaluated using six reference strains and nine clinical isolates. RESULTS: Using optimized conditions, all carbapenemase-positive controls yielded predicted amplicon sizes and confirmed the specificity of the primers in single and multiplex PCR. CONCLUSIONS: We have reported here a reliable method, composed of single and multiplex PCR assays, for screening all clinically known carbapenemases. Primers tested in silico and in vitro may distinguish carbapenem-resistant Enterobacteriaceae and could assist in combating the spread of carbapenem resistance in Enterobacteriaceae.
Citace poskytuje Crossref.org
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