Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci by PCR

Following the identification of two clinical isolates of vancomycin-resistant enterococci (VRE) from intensive care unit (ICU) patients, a surveillance programme detected that six of eight ICU patients were colonised by VRE. Standard epidemic control measures were instituted in the ICU. During a 16-month period, 13 (2.5%) of 509 ICU patients had VRE-positive swabs upon admission, and 43 (8.7%) of 496 VRE-negative patients were colonised by VRE in the ICU. Patients who acquired VRE in the ICU had a longer ICU stay (p < 0.0001). No other statistically significant differences were demonstrated. Two patients had documented infection (infection/colonisation index, 3.6%; overall VRE infection frequency, 0.4%), but both recovered and were discharged. VRE colonisation did not increase the mortality rate. Automated ribotyping identified three clusters containing, respectively, the first 52 Enterococcus faecium isolates, two Enterococcus faecalis isolates, and two further isolates of E. faecium. Multilocus sequence typing demonstrated that two E. faecium isolates representative of the two ribotypes belonged to sequence types 78 and 18, and that these two isolates belonged to the epidemic lineage C1, which includes isolates with a wide circulation in northern Italy. The outbreak was controlled by continuous implementation of the infection control programme, and by the opening of a new unit with an improved structural design and hand-washing facilities.

Section: Genotypic Resistancementioning

confidence: 99%

Outbreak of vancomycin-resistant Enterococcus spp. in an Italian general intensive care unit

Peta

Carretto

Barbarini

et al. 2006

“…DNA extraction was performed with the Instagene Matrix commercial kit (BioRad, Hercules, Ca, USA) according to the manufacturer's instructions. Primers, reaction mixtures and PCR conditions followed published studies for the detection of vanA [15], vunB [16], vanCl [17] and vanC2 [18] genes. Enterococcus fuecium U2A1 [19], E.fecalis SF-299 (provided by George M. Eliopoulos), E .…”

Section: Methodsmentioning

confidence: 99%

MIC distribution and inoculum effect of LY333328: a study of vancomycin-susceptible and VanA-type and VanC-type enterococci obtained from intensive care unit patient surveillance cultures

Mir

Sánchez

Baquero

1999

OBJECTIVE: To determine the in vitro activity and inoculum effect of LY333328, a semisynthetic glycopeptide, against vancomycin-susceptible and vancomycin-resistant enterococcal isolates. METHODS: One hundred and seventy-six enterococcal isolates (117 vancomycin-susceptible, 29 VanA-type and 30 VanC-type isolates) obtained from surveillance cultures of 139 intensive care unit patients were studied by the standard agar dilution method. Vancomycin resistance determinants were characterized by PCR. RESULTS: The activity of LY333328 was comparable (MIC range, 0.1-2 mg/L) to those of vancomycin (0.1-4 mg/L) and teicoplanin (0.06-1 mg/L) for vancomycin-susceptible isolates. LY333328 was more active (0.1-8 mg/L) than vancomycin (256 to >1024 mg/L) and teicoplanin (32-512 mg/L) against VanA-type isolates, and similar (0.2-1 mg/L) to teicoplanin (0.1-0.5 mg/L) against VanC-type isolates. The MIC distribution of LY333328 displayed a narrower range than that of vancomycin, with no clear distinction between susceptible and resistant populations. The increment in the inoculum size, from 104 to 106 CFU/spot, of susceptible isolates increased the MIC values of LY333328, vancomycin and teicoplanin by factors of 11.4, 1.6 and 3.8, respectively. The corresponding factors for LY333328 for VanA-type and VanC-type isolates were 3.5 and 6.4, respectively. CONCLUSIONS: LY333328 displays an excellent in vitro activity against vancomycin-susceptible and -resistant enterococci. Nevertheless, the inoculum size used in susceptibility tests should be carefully controlled.

“…One isolate of glycopeptide-susceptible E. faecalis was included in each PFGE run as a control. Vancomycin resistance genotypes were determined by PCR with vanA-and vanB-specific primers [16,17] selected from published gene sequences.…”

Section: Clin Microbiol Infect 2004; 10: 260-262mentioning

confidence: 99%

Inter-hospital dissemination of glycopeptide-resistant Enterococcus faecalis in Brazil

Cordeiro

Silbert

Reis

et al. 2004

The antimicrobial susceptibility patterns of 73 glycopeptide-resistant Enterococcus faecalis isolates from nine hospitals in Brazil were analysed by the disk diffusion method and Etests. Isolates were typed by pulsed-field gel electrophoresis (PFGE), and vancomycin resistance genes were detected by PCR. The isolates shared a single major PFGE pattern, with six subtypes, and all were positive for vanA. These results indicate the occurrence of inter-hospital dissemination of glycopeptide-resistant E. faecalis in São Paulo, and raise concerns about the rapid dissemination of this pathogen throughout Brazil.