scholarly journals 97. Assessment of the accuracy of direct antimicrobial susceptibility testing from positive blood cultures in pediatric patients and its utility as an antimicrobial stewardship tool

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S63-S64
Author(s):  
Timothy Savage ◽  
Shun Rao ◽  
Jill Joerger ◽  
Al Ozonoff ◽  
Alexander McAdam ◽  
...  

Abstract Background Hospitalized pediatric patients with bacteremia receive broad-spectrum antibiotics while organism identification and antibiotic susceptibilities are pending. Direct susceptibility testing (DST) using unstandardized Kirby-Bauer disk diffusion provides early results before standardized (final) antimicrobial susceptibility testing (AST) is available. The accuracy of DST in comparison with AST has been poorly studied. If DST is highly accurate, it could facilitate earlier de-escalation of antibiotics. Methods Retrospective cohort study of all positive blood cultures at Boston Children’s Hospital between January 1, 2017 and October 20, 2019. Isolates were eligible for inclusion if a DST result was available. Isolates were excluded if more than one organism grew from a blood culture or if a patient had repeat blood cultures positive for the same organism within 14 days. Patient characteristics and antibiotic orders were identified via a local data warehouse. Positive and negative predictive value (PPV: accuracy of susceptibility on DST in identifying susceptibility on AST result; NPV: accuracy of DST in identifying non-susceptibility on AST result) and 95% confidence interval were calculated for each bug-drug combination. Antibiotic Spectrum Index was evaluated at 3 time points to assess change in antibiotic prescribing after availability of DST and AST results. Results 496 patients (median age: 51 months, IQR: 7–165 months) with 603 positive blood cultures were included in the final analysis. PPV of DST was ≥96%for most organism-antibiotic pairs (Table). NPV of DST varied substantially across organism-antibiotic pairs and was frequently lower than PPV. The proportion of patients with more narrow spectrum antibiotic orders increased after the DST result and again after the AST result (Figure). Table Test performance of direct susceptibility testing (DST) of clinical isolates from patients with bacteremia to identify organism susceptibility to the noted antibiotic. Positive predictive value measures the ability of a susceptible DST result to predict a susceptible AST result. Negative predict value measures the ability of an intermediate or resistant DST result to predict an intermediate or resistant AST result. *Includes Staphylococcus capitis, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus pettenkoferi, and Staphylococcus warneri. ‡Includes Citrobacter freundii complex, Citrobacter koseri, Enterobacter aerogenes, Enterobacter cloacae complex, Escherichia coli, Escherichia vulneris, Klebsiella oxytoca, Klebseilla pneumoniae, Klebsiella variicola, Pantoea species, Proteus mirabilis, Salmonella species, Salmonella typhi, Serratia liquefaciens, and Serratia marcescens. Figure Distribution of spectrum of antibiotics ordered to treat bacteremia in pediatric patients. Antibiotic spectrum, as measured by the antibiotic spectrum index, is represented in this Sankey diagram on the Y-axis. The height of each group indicates the proportion of patients with an aggregate antibiotic spectrum falling in that category at three time points. The ASI of empiric antibiotics was measured just before the DST result to provide time for a treating physician to settle on an empiric regimen. The ASI with the DST result was measured just before the ASI result. The ASI with the AST result was measured 24 hours after the AST result was available. Green bars indicate the proportion of patients within a group whose ASI remained unchanged between time points. Blue bars indicate the proportion of patients within a group whose ASI decreased, and red bars indicate the proportion of patients whose ASI increased between time points. Conclusion DST is highly accurate at identifying susceptibility to antibiotics for many bug-drug combinations in pediatric blood culture isolates, but its ability to identify non-susceptibility is less robust. The observed spectrum of prescribed antibiotics was narrower after DST results, suggesting some clinicians may be using the result to de-escalate therapy. DST may be a useful low-cost tool for antimicrobial stewardship. Disclosures All Authors: No reported disclosures

2017 ◽  
Vol 55 (7) ◽  
pp. 2116-2126 ◽  
Author(s):  
Matthias Marschal ◽  
Johanna Bachmaier ◽  
Ingo Autenrieth ◽  
Philipp Oberhettinger ◽  
Matthias Willmann ◽  
...  

ABSTRACT Bloodstream infections (BSI) are an important cause of morbidity and mortality. Increasing rates of antimicrobial-resistant pathogens limit treatment options, prompting an empirical use of broad-range antibiotics. Fast and reliable diagnostic tools are needed to provide adequate therapy in a timely manner and to enable a de-escalation of treatment. The Accelerate Pheno system (Accelerate Diagnostics, USA) is a fully automated test system that performs both identification and antimicrobial susceptibility testing (AST) directly from positive blood cultures within approximately 7 h. In total, 115 episodes of BSI with Gram-negative bacteria were included in our study and compared to conventional culture-based methods. The Accelerate Pheno system correctly identified 88.7% (102 of 115) of all BSI episodes and 97.1% (102 of 105) of isolates that are covered by the system's identification panel. The Accelerate Pheno system generated an AST result for 91.3% (95 of 104) samples in which the Accelerate Pheno system identified a Gram-negative pathogen. The overall category agreement between the Accelerate Pheno system and culture-based AST was 96.4%, the rates for minor discrepancies 1.4%, major discrepancies 2.3%, and very major discrepancies 1.0%. Of note, ceftriaxone, piperacillin-tazobactam, and carbapenem resistance was correctly detected in blood culture specimens with extended-spectrum beta-lactamase-producing Escherichia coli ( n = 7) and multidrug-resistant Pseudomonas aeruginosa ( n = 3) strains. The utilization of the Accelerate Pheno system reduced the time to result for identification by 27.49 h ( P < 0.0001) and for AST by 40.39 h ( P < 0.0001) compared to culture-based methods in our laboratory setting. In conclusion, the Accelerate Pheno system provided fast, reliable results while significantly improving turnaround time in blood culture diagnostics of Gram-negative BSI.


Author(s):  
Timothy J. Savage ◽  
Shun Rao ◽  
Jill Joerger ◽  
Al Ozonoff ◽  
Alexander J. McAdam ◽  
...  

Background Accurate and early susceptibility results could reduce overuse of broad-spectrum antibiotics for empiric treatment of bacteremia. Direct disk diffusion testing (dDD) using non-standardized inocula directly from blood cultures could facilitate earlier narrowing of antibiotics. Methods To determine the predictive value of dDD compared with standardized antimicrobial susceptibility testing (AST), we performed a retrospective cohort study of 582 blood cultures from 495 pediatric patients with bacteremia. Positive and negative predictive value (PPV: number of isolates susceptible by both dDD and AST divided by the total number of isolates susceptible by dDD; NPV: number of isolates not susceptible [either intermediate or resistant] by both dDD and AST divided by the total number of isolates not susceptible by dDD), sensitivity, specificity, and 95% confidence interval were calculated for each bacterium-antibiotic combination. We evaluated the Antibiotic Spectrum Index of prescribed antibiotics to assess change in antibiotic prescribing after availability of Gram stain, dDD, and AST results. Results dDD results were available a median of 21 hours before AST results. dDD had PPVs of ≥96% for most organism-antibiotic pairs, including 100% (CI 96-100%) for Staphylococcus aureus and oxacillin and 99% (CI 93%-100%) for Enterobacterales and ceftriaxone. NPVs of dDD were variable and frequently lower than PPV. Very major errors and major errors occurred in 31/5454 (0.6%) and 231/5454 (4.2%) organism-antibiotic combinations, respectively. Antibiotics were narrowed in 30% of cases after dDD result and a further 25% of cases after AST result. Conclusions dDD is highly predictive of susceptibility for many common organism-antibiotic combinations and provides actionable information one day earlier than standard susceptibility approaches. dDD has the potential to facilitate earlier de-escalation to narrow-spectrum antibiotic treatment.


Author(s):  
Måns Ullberg ◽  
Volkan Özenci

Abstract Rapid identification and antimicrobial susceptibility testing remain a crucial step for early efficient therapy of bloodstream infections. Traditional methods require turnaround times of at least 2 days, while rapid procedures are often associated with extended hands-on time. The Accelerate Pheno™ System provides microbial identification results within 90 min and susceptibility data in approximately 7 h directly from positive blood cultures with only few minutes of hands-on time. The aim of this study was, therefore, to evaluate the performance of the Accelerate Pheno™ System in identification and antimicrobial susceptibility testing of both Gram-positive and Gram-negative bacteria directly from clinical blood culture samples. We analyzed 108 and 67 blood culture bottles using the Accelerate PhenoTest™ BC kit with software version v1.0 and the FDA-cleared version v1.2, respectively. Reliable identification was achieved for Enterobacteriaceae, staphylococci, and enterococci, with 76/80 (95%), 42/46 (91%), and 10/11 (91%) correct identifications. Limitations were observed in the identification of streptococci, including Streptococcus pneumoniae and Streptococcus pyogenes, and coagulase-negative staphylococci. Antimicrobial susceptibility results for Enterobacteriaceae, for amikacin, ertapenem, ciprofloxacin, gentamicin, meropenem, and piperacillin-tazobactam ranged between 86 and 100% categorical agreement. Using v1.2, results for ceftazidime showed 100% concordance with the reference method. For staphylococci, the overall performance reached 92% using v1.2. Qualitative tests for detection of methicillin or macrolide-lincosamide-streptogramin B (MLSB) resistance caused major and very major errors for isolates. Overall, the present data show that the Accelerate Pheno™ system can, in combination with Gram stain, be used as a rapid complementation to standard microbial diagnosis of bloodstream infections.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Suzanne Putney ◽  
Andrew H. Theiss ◽  
Nitin K. Rajan ◽  
Eszter Deak ◽  
Creighton Buie ◽  
...  

AbstractA key predictor of morbidity and mortality for patients with a bloodstream infection is time to appropriate antimicrobial therapy. Accelerating antimicrobial susceptibility testing from positive blood cultures is therefore key to improving patient outcomes, yet traditional laboratory approaches can require 2–4 days for actionable results. The eQUANT—a novel instrument utilizing electrical biosensors—produces a standardized inoculum equivalent to a 0.5 McFarland directly from positive blood cultures. This proof-of-concept study demonstrates that eQUANT inocula prepared from clinically significant species of Enterobacterales were comparable to 0.5 McF inocula generated from bacterial colonies in both CFU/ml concentration and performance in antimicrobial susceptibility testing, with ≥ 95% essential and categorical agreement for VITEK2 and disk diffusion. The eQUANT, combined with a rapid, direct from positive blood culture identification technique, can allow the clinical laboratory to begin antimicrobial susceptibility testing using a standardized inoculum approximately 2–3 h after a blood culture flags positive. This has the potential to improve clinical practice by accelerating conventional antimicrobial susceptibility testing and the resulting targeted antibiotic therapy.


2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S387-S387
Author(s):  
Nadim Khalil ◽  
Eleanor Powell ◽  
Joel Mortensen

Abstract Background Infections secondary to pathogens resistant to third-generation cephalosporins (3GC), such as extended-spectrum (ESBL) and AmpC β-lactamase (AmpC) producing Enterobacteriaceae, are increasing. Currently, there are no recommendations regarding identification of AmpC in Citrobacter, Enterobacter, Morganella and Serratia spp. (CEMS organisms). This study’s aim was to increase the detection of AmpC and ESBL producing Enterobacteriaceae in blood cultures from pediatric population by combining genotypic with phenotypic antimicrobial susceptibility testing (AST). Methods All first time Enterobacteriaceae isolates recovered from blood cultures of pediatric patients at CCHMC between January 2017 and December 2018 were evaluated. The Check-MDR CT103XL assay was used to determine the presence of AmpC and ESBL. AST was performed using the Vitek 2 platform. Phenotypic ESBL resistance was defined by resistant to either ceftriaxone of ceftazidime using CLSI breakpoints. Combined cefoxitin resistance with ceftriaxone or ceftazidime resistance was used to define phenotypic AmpC (EUCAST standards). Results There were 170 isolates, from 147 patients, with 21 (12.4%) AmpC and 18 (10.6%) ESBL genes detected. Using AST, 11 (6.5%) and 26 (15.3%) isolates met AmpC and ESBL phenotypic criteria respectively. However, 14 of the isolates with AmpC genes detected and 2 of isolates with ESBL genes detected failed to meet phenotypic criteria. In addition, 4 (19%) of 21 AmpC isolates were susceptible to cefoxitin and 3GC while both E. coli and S. marcescens genotypic ESBL isolates were susceptible to 3GC. Number of AmpC- and ESBL- Producing Isolates Detected Using Phenotypic Method a,b, Genotypic Method and Combined Testing Conclusion We identified 16 (9.4%) isolates with resistance genes detected but that failed to meet phenotypic criteria. Without molecular testing, patients with these isolates may have been treated with 3GC which could have resulted in treatment failure. The addition of genotypic testing to AST improved the identification of AmpC and ESBL organisms and provided clinically relevant data to guide treatment of resistant organisms. Combined testing is also beneficial for infection control and epidemiological purposes. Disclosures All Authors: No reported disclosures


2020 ◽  
Vol 75 (11) ◽  
pp. 3218-3229
Author(s):  
Stefano Mancini ◽  
Elias Bodendoerfer ◽  
Natalia Kolensnik-Goldmann ◽  
Sebastian Herren ◽  
Kim Röthlin ◽  
...  

Abstract Background Rapid antimicrobial susceptibility testing (RAST) of bacteria causing bloodstream infections is critical for implementation of appropriate antibiotic regimens. Objectives We have established a procedure to prepare standardized bacterial inocula for Enterobacterales-containing clinical blood cultures and assessed antimicrobial susceptibility testing (AST) data generated with the WASPLabTM automated reading system. Methods A total of 258 blood cultures containing Enterobacterales were examined. Bacteria were enumerated by flow cytometry using the UF-4000 system and adjusted to an inoculum of 106 cfu/mL. Disc diffusion plates were automatically streaked, incubated for 6, 8 and 18 h and imaged using the fully automated WASPLabTM system. Growth inhibition zones were compared with those obtained with inocula prepared from primary subcultures following the EUCAST standard method. Due to time-dependent variations of the inhibition zone diameters, early AST readings were interpreted using time-adjusted tentative breakpoints and areas of technical uncertainty. Results and conclusions Inhibition zones obtained after 18 h incubation using an inoculum of 106 cfu/mL prepared directly from blood cultures were highly concordant with those of the EUCAST standard method based on primary subcultures, with categorical agreement (CA) of 95.8%. After 6 and 8 h incubation, 89.5% and 93.0% of the isolates produced interpretable results, respectively, with CA of &gt;98.5% and very low numbers of clinical categorization errors for both the 6 h and 8 h readings. Overall, with the standardized and automated RAST method, consistent AST data from blood cultures containing Enterobacterales can be generated after 6–8 h of incubation and subsequently confirmed by standard reading of the same plate after 18 h.


2021 ◽  
Vol 12 ◽  
Author(s):  
Min Cao ◽  
Lin Huang ◽  
Yanyan Hu ◽  
Yinfei Fang ◽  
Rong Zhang ◽  
...  

Bloodstream infections (BSI) are associated with high morbidity and mortality and remain a leading cause of death. Blood culture (BC) including the identification and the antimicrobial susceptibility testing of the causative microorganisms should be performed as soon as possible. In this study, we developed an in-house rapid antimicrobial susceptibility testing (rAST) protocol for positive BC. First, the rAST was performed in the simulated positive BC of standard strains (Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, and Pseudomonas aeruginosa ATCC 27853) at three different times to assess the reproducibility and operability by dispensing four drops of BC broth onto a Mueller–Hinton agar plate after a positive signal. Furthermore, the rAST was performed in clinical positive BCs. The results of rAST at 4, 6, 8, and 18 h of incubation were compared with results of the standard 16- to 20-h disk diffusion method, and the preliminary breakpoints of the rAST method were established according to the inhibition diameter of sensitive strains and resistant strains. Finally, the rAST was performed in the simulated positive BC of clinical strains to evaluate the availability of the preliminary breakpoints. The rAST results of standard strains were distributed evenly at three different times. Among the 202 clinical strains used to establish the preliminary breakpoints, the number of zone diameters that could be read and interpreted (60, 87, 98, and 100%) increased with incubation time (4, 6, 8, and 18 h), and the categorical agreement was acceptable, with total error rates of 3.0, 2.3, 2.1, and 1.3% at 4, 6, 8, and 18 h of incubation, respectively. In conclusion, the in-house rAST protocol for positive BC can be implemented in routine laboratories. It provides reliable antimicrobial susceptibility testing results for BSI pathogens after 4–6 h of incubation.


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