Reverse transcription polymerase chain reaction method for the detection of glycopeptide resistance in enterococci
Articolo
Data di Pubblicazione:
1999
Citazione:
Reverse transcription polymerase chain reaction method for the detection of glycopeptide resistance in enterococci / O. Privitera, F. Sisto, V. Giuffrida, M. Puntorieri, C. Cascone, I. Di Silvestro, G. Rappazzo, S. Stefani. - In: JOURNAL OF MICROBIOLOGICAL METHODS. - ISSN 0167-7012. - 35:2(1999 Mar), pp. 95-100.
Abstract:
In this work we have developed reverse transcription polymerase chain reaction (RT-PCR) methods for detecting specific mRNA from enterococci, particularlyvanA andvanB genes, responsible for glycopeptide resistance in this genus. mRNA from the two genes was detected immediately after RNA extraction of a midlog phase culture, determined by growth rate analysis. Because of the short half-life associated with many bacterial RNA species (1.5–2 min), time is an important factor in obtaining RNA of good yield and high purity. Our results showed that: (i) the transcription of mRNA tovanA ligase in enterococci showing Van A phenotype happens only after induction with both vancomycin and teicoplanin; (ii) the transcription of mRNA related tovanB ligase happens only in the presence of vancomycin and (iii) there was no transcription of mRNA in the two strains positive tovanA gene after PCR experiments.
RT-PCR methodology can have numerous applications in microbiology for studying gene expression in isolated bacteria and also in nonculturable cells in environmental samples, for studies of mechanisms and/or as an indicator of viability in bacterial communities.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
RT-PCR ; Enterococcus ; Glycopeptide ; Mechanisms of resistance
Elenco autori:
O. Privitera, F. Sisto, V. Giuffrida, M. Puntorieri, C. Cascone, I. Di Silvestro, G. Rappazzo, S. Stefani
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