Rapid detection of vancomycin-non-susceptible Staphylococcus aureus using the spiral gradient endpoint technique.

OBJECTIVES: Several methods have been introduced for detection of vancomycin-non-susceptible Staphylococcus aureus [heterogeneous vancomycin-intermediate S. aureus (hVISA) and vancomycin-intermediate S. aureus (VISA)]. However, the limitations of these methods can delay appropriate therapy for the patient. This study evaluated the spiral gradient endpoint (SGE) technique for detection of hVISA/VISA. METHODS: The SGE method was evaluated for intra-batch, inter-batch and inter-observer reproducibility in comparison with MICs determined by agar dilution. Three media, Mueller-Hinton agar, brain heart infusion agar and brain heart infusion agar with 5% glucose, were evaluated. The SGE method was compared with agar dilution for correlation of MIC and susceptibility category using control strains, clinical isolates and induced vancomycin-non-susceptible strains. RESULTS: The SGE method had good reproducibility and there was excellent correlation of MICs generated by SGE using brain heart infusion agar with those by agar dilution (r(2) =0.950), with no difference in resistance categories generated by the two methods. All VISA isolates were correctly identified and the method allowed easy identification of hVISA by means of the trailing endpoint. CONCLUSIONS: SGE offers a simple, rapid and cost-effective alternative method for the detection of hVISA/VISA for the routine laboratory. Early recognition of vancomycin-non-susceptible strains can allow the change to appropriate antibiotics, resulting in potentially better patient outcomes.

Regional differences in temporal incidence of Clostridium difficile infection: a systematic review and meta-analysis.

BACKGROUND: Previous decades have witnessed a change in the epidemiology of Clostridium difficile infections. This study aimed to determine temporal trends in the incidence of C difficile infection across geographic regions. METHODS: An initial search of the relevant literature was conducted from date inception to October 2018 without language restriction. We estimated the pooled incidences using logit transformation, weighted by inverse variance. The Joinpoint Regression Analysis Program was used to explore its temporal trend. RESULTS: Globally, the estimated incidence of C difficile infection increased from 6.60 per 10,000 patient-days in 1997 to 13.8 per 10,000 patient-days in 2004. Thereafter, a significant downward trend was observed, at -8.75% annually until 2015. From 2005 to 2015, the incidences in most European countries decreased at a rate between 1.97% and 4.11% per annum, except in France, where an increasing incidence was observed (ß = 0.16; P < .001). The incidences have stabilized in North America over the same period; however, in Asia, the incidence increased significantly from 2006 to 2014 (annualized percentage change = 14.4%; P < .001). The increase was greatest in Western Asian countries, including Turkey and Israel (ß > 0.10; P < .004). CONCLUSIONS: This study revealed rapid changes in the incidence of C difficile infection. This meta-analysis should inform the allocation of resources for controlling C difficile infection and future surveillance efforts in countries where epidemiologic information on C difficile infection remains sparse.

Reduced vancomycin susceptibility in porcine ST9 MRSA isolates.

Porcine strains of livestock-associated methicillin resistant Staphylococcus aureus (LA-MRSA) have been recognized in many countries and have been shown to be able to cause human infection. Resistance to non-beta lactam antibiotics has been reported but non-susceptibility to vancomycin, which is known to occur in human MRSA, has so far not been observed in LA-MRSA. Such resistance is typically fairly low level involving changes in the cell wall thickness. The development of resistance is usually preceded by presence of a sub-population having an increased MIC, which is selected for by exposure to vancomycin. This study investigated vancomycin susceptibility of one hundred porcine MRSA isolates using three MIC methods including spiral gradient endpoint (SGE) technique which allows visualization of more resistant sub-populations. SGE revealed 16 strains with an MIC above 2.0 mg/L, of which 14 were determined to have MIC 4 mg/L by agar dilution (AD). SGE revealed a further two isolates with MIC < 2 mg/L had a sub-population >2 mg/L. In addition, trailing endpoints not reaching resistance were present in 26 isolates with MIC < 2 mg/L. Sequencing of the genes of the VraSR/GraSR two component systems of ten of the resistant strains for comparison with susceptible strains revealed changes, including the presence of stop codons, in vraS and graR, but these were not consistent in all isolates. Other genetic changes may contribute to vancomycin non-susceptibility and require investigation. As failure to respond to treatment has been reported in clinical isolates with MIC > 1.5 mg/L, the presence of vancomycin non-susceptibility in porcine isolates is of concern and further monitoring of LA-MRSA is essential.