Identification by mass spectrometry and automated susceptibility testing from positive bottles: a simple, rapid, and standardized approach to reduce the turnaround time in the management of blood cultures.

BACKGROUND: Speeding up identification and antimicrobial susceptibility testing (AST) is of foremost importance in the management of blood cultures. Here, we describe a simple, rapid, and standardized approach based on a very short-term incubation on solid medium from positive blood cultures followed by MALDI-TOF mass spectrometry identification and automated AST. The aim of the study was to evaluate the impact in the laboratory practice of this new procedure with respect to that previously used (standard method) by comparing TAT and cumulative percentage of final reports to clinicians. RESULTS: Compared with the standard method, the new procedure gave correct organism identification at genus or species level in 98.4% of monomicrobial samples. AST resulted in 97.7% essential agreement and 98.1% categorical agreement, with 0.9% minor errors, 1.0% major error, and 1.5% very major errors. The mean turnaround time to identification and AST was 61.4 h by using the new method compared to 83.1 h by using standard procedure. Concerning cumulative percentages of final reports, approximately a third of results were available at 48 h from the check-in of the sample when using the new procedure, whereas no final reports were ready at the same time with the standard method. CONCLUSIONS: The new procedure allows faster and reliable results using a simple and standardized approach. Thus, it represents an important tool for a more rapid management of blood cultures when molecular methods are not available in the laboratory.

Evaluation of brilliance CRE agar for the detection of carbapenem-resistant gram-negative bacteria.

The aim of this work was to evaluate the performance of the new chromogenic medium BrillianceTM CREAgar (Thermo Fisher Scientific) for determining the limit of detection of carbapenem-resistant enterobacteria (CRE). A total of 70 clinical isolates were studied. Of these, 30 were well-characterized CRE, including Klebsiella pneumoniae strains producing KPC-, VIM-, and OXA-type enzymes, VIM-positive Enterobacter cloacae and Escherichia coli, NDM-positive E. coli, and enterobacterial isolates characterized by porin loss associated with ESBL production or AmpC hyperproduction. Ten carbapenem-resistant non-fermentative isolates were also included as well as 30 carbapenem-susceptible isolates. Carbapenem-resistant strains were inoculated at three different concentrations onto Brilliance CRE Agar (from 1.5x101 CFU/ml up to 1.5x104 CFU/ml) whereas carbapenem-susceptible isolates were inoculated at a concentration of 1.5x102 CFU/ml. The medium sustained the growth of carbapenem-resistant isolates, showing detection limits from 1.5x101 CFU/ml (in 31/40 cases) to 1.5x104 CFU/ml. No growth was observed with carbapenem-sensitive control strains. Our results indicate that the Brilliance CRE Agar allows the growth of carbapenem-resistant isolates with low detection limits and could represent a useful screening medium for both enterobacteria and non-fermentative Gram-negative strains resistant to carbapenems.