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Background: Patient mortality is significantly reduced by rapid identification of bacteria from sterile sites. MALDI-TOF can identify bacteria directly from positive blood cultures and multiple sample preparation methods are available. We evaluated three sample preparation methods and two MALDI-TOF score cut-off values. Positive blood culture bottles with organisms present in Gram stains were prospectively analysed by MALDI-TOF. Three lysis reagents (Saponin, SDS, and SepsiTyper lysis bufer) were applied to each positive culture followed by centrifugation, washing and protein extraction steps. Methods were compared using the McNemar test and 16S rDNA sequencing was used to assess discordant results. Results: In 144 monomicrobial cultures, using ≥2.000 as the cut-off value, species level identifications were obtained from 69/144 (48%) samples using Saponin, 86/144 (60%) using SDS, and 91/144 (63%) using SepsiTyper. The difference between SDS and SepsiTyper was not statistically significant (P = 0.228). Differences between Saponin and the other two reagents were significant (P < 0.01). Using ≥1.700 plus top three results matching as the cut-off value, species level identifications were obtained from 100/144 (69%) samples using Saponin, 103/144 (72%) using SDS, and 106/144 (74%) using SepsiTyper and there was no statistical difference between the methods. No true discordances between culture and direct MALDI-TOF identification were observed in monomicrobial cultures. In 32 polymicrobial cultures, MALDI-TOF identified one organism in 34–75% of samples depending on the method. Conclusions: This study demonstrates two inexpensive in-house detergent lysis methods are non-inferior to a commercial kit for analysis of positive blood cultures by direct MALDI-TOF in a clinical diagnostic microbiology laboratory. [ABSTRACT FROM AUTHOR]

Background: Faster identification of bacterial isolates from blood cultures can enable earlier clinical intervention for patients with sepsis. We evaluated the clinical impact of direct identification of micro-organisms from positive blood cultures using MALDI-ToF. Method: Positive blood cultures with organisms seen on Gram stain were included over a four week period. For each patient case, comparison was made between the clinical advice given on day one with only a Gram stain result, and the follow up advice given on day two with the benefit of organism identification. Culture results were then compared with direct MALDI-ToF identification. Results: For 73 of 115 cases (63.5%), direct organism identification was obtained by MALDI-ToF. Of those 73, 70 (95.5%) had a result concordant with that of the plate culture. In 28 of the 115 cases (24.3%) direct MALDI-ToF identification on day one would have had a clear clinical benefit. In 11 cases it would have helped to identify the potential source of bacteraemia. In 11 cases it would have indicated a different antibiotic regimen on day one, with five patients receiving appropriate antibiotics 24 hours earlier. For 14 cases the blood culture isolate could have been designated as unlikely to be clinically significant. Conclusion: We have demonstrated that organism identification on day one of blood culture positivity can have a direct clinical impact. Faster identification using MALDI-ToF assists the clinician in assessing the significance of a blood culture isolate on day one. It can allow earlier appropriate choice of antimicrobial agent, even in the absence of susceptibility testing, and help narrow down the potential source of infection providing a focus for further investigation in a more timely way than conventional techniques alone. [ABSTRACT FROM AUTHOR]

Background Antimicrobial resistance (AMR) is recognized as one of the most significant threats to human health. Local and regional AMR surveillance enables the monitoring of temporal changes in susceptibility to antibiotics and can provide prescribing guidance to healthcare providers to improve patient management and help slow the spread of antibiotic resistance in the community. There is currently a paucity of routine community-level AMR surveillance information. Methods The HPA in England sponsored the development of an AMR surveillance system (AmSurv) to collate local laboratory reports. In the West Midlands region of England, routine reporting of AMR data has been established via the AmSurv system from all diagnostic microbiology laboratories. The HPA Regional Epidemiology Unit developed a web-enabled database application (AmWeb) to provide microbiologists, pharmacists and other stakeholders with timely access to AMR data using user-configurable reporting tools. Results AmWeb was launched in the West Midlands in January 2012 and is used by microbiologists and pharmacists to monitor resistance profiles, perform local benchmarking and compile data for infection control reports. AmWeb is now being rolled out to all English regions. Conclusions It is expected that AmWeb will become a valuable tool for monitoring the threat from newly emerging or currently circulating resistant organisms and helping antibiotic prescribers to select the best treatment options for their patients. [ABSTRACT FROM AUTHOR]