scholarly journals Evaluation of digital dispense-assisted broth microdilution antimicrobial susceptibility testing for Pseudomonas aeruginosa isolates

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Shawn T. Clark ◽  
Patrick J. Stapleton ◽  
Pauline W. Wang ◽  
Yvonne C. W. Yau ◽  
Valerie J. Waters ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antimicrobial Susceptibility ◽  
Inkjet Printing ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Broth Microdilution ◽  
Standard Methods ◽  
Error Measures ◽  
Inkjet Printing Technology ◽  
Variable Performance

AbstractAntimicrobial susceptibility testing (AST) is essential for detecting resistance in Pseudomonas aeruginosa and other bacterial pathogens. Here we evaluated the performance of broth microdilution (BMD) panels created using a semi-automated liquid handler, the D300e Digital Dispenser (Tecan Group Ltd., CH) that relies on inkjet printing technology. Microtitre panels (96-well) containing nine twofold dilutions of 12 antimicrobials from five classes (β-lactams, β-lactam/β-lactamase inhibitors, aminoglycosides, fluoroquinolones, polymyxins) were prepared in parallel using the D300e Digital Dispenser and standard methods described by CLSI/ISO. To assess performance, panels were challenged with three well characterized quality control organisms and 100 clinical P. aeruginosa isolates. Traditional agreement and error measures were used for evaluation. Essential (EA) and categorical (CA) agreements were 92.7% and 98.0% respectively for P. aeruginosa isolates with evaluable on-scale results. The majority of minor errors that fell outside acceptable EA parameters (≥ ± 1 dilution, 1.9%) were seen with aztreonam (5%) and ceftazidime (4%), however all antimicrobials displayed acceptable performance in this situation. Differences in MIC were often log2 dilution lower for D300e dispensed panels. Major and very major errors were noted for aztreonam (2.6%) and cefepime (1.7%) respectively. The variable performance of D300e panels suggests that further testing is required to confirm their diagnostic utility for P. aeruginosa.

scholarly journals Multicenter Evaluation of Colistin Broth Disk Elution and Colistin Agar Test: a Report from the Clinical and Laboratory Standards Institute

2019 ◽  
Vol 57 (11) ◽  
Author(s):  
Romney M. Humphries ◽  
Daniel A. Green ◽  
Audrey N. Schuetz ◽  
Yehudit Bergman ◽  
Shawna Lewis ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Polymyxin B ◽  
Antimicrobial Susceptibility Testing ◽  
Broth Microdilution ◽  
Content Type ◽  
Clinical Laboratories

ABSTRACT Susceptibility testing of the polymyxins (colistin and polymyxin B) is challenging for clinical laboratories. The Clinical and Laboratory Standards Institute (CLSI) Antimicrobial Susceptibility Testing Subcommittee evaluated two methods to enable accurate testing of these agents. These methods were a colistin broth disk elution (CBDE) and a colistin agar test (CAT), the latter of which was evaluated using two inoculum volumes, 1 μl (CAT-1) and 10 μl (CAT-10). The methods were evaluated using a collection of 270 isolates of Enterobacterales, 122 Pseudomonas aeruginosa isolates, and 106 Acinetobacter spp. isolates. Overall, 94.4% of CBDE results were in essential agreement and 97.9% in categorical agreement (CA) with reference broth microdilution MICs. Nine very major errors (VME; 3.2%) and 3 major errors (ME; 0.9%) were observed. With the CBDE, 98.6% CA was observed for Enterobacterales (2.5% VME, 0% ME), 99.3% CA was observed for P. aeruginosa (0% VME, 0.7% ME), and 93.1% CA was observed for Acinetobacter spp. (5.6% VME, 3.3% ME). Overall, CA was 94.9% with 6.8% VME using CAT-1 and improved to 98.3% with 3.9% VME using CAT-10. No ME were observed using either CAT-1 or CAT-10. Using the CAT-1/CAT-10, the CA observed was 99.4%/99.7% for Enterobacterales (1%/0.5% VME), 98.7%/100% for P. aeruginosa (8.3%/0% VME), and 88.5%/92.3% for Acinetobacter spp. (21.4%/14.3% VME). Based on these data, the CLSI antimicrobial susceptibility testing (AST) subcommittee endorsed the CBDE and CAT-10 methods for colistin testing of Enterobacterales and P. aeruginosa.

scholarly journals Performance of Vitek 2 for Antimicrobial Susceptibility Testing of Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia with Vitek 2 (2009 FDA) and CLSI M100S 26th Edition Breakpoints

2016 ◽  
Vol 55 (2) ◽  
pp. 450-456 ◽  
Author(s):  
April M. Bobenchik ◽  
Eszter Deak ◽  
Janet A. Hindler ◽  
Carmen L. Charlton ◽  
Romney M. Humphries
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Acinetobacter Baumannii ◽  
Antimicrobial Susceptibility ◽  
Stenotrophomonas Maltophilia ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Broth Microdilution ◽  
Content Type ◽  
Vitek 2 ◽  
Improved Performance

ABSTRACTThe performances of Vitek 2 AST-GN69 and AST-XN06 cards were compared to Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution (BMD) for 99 isolates ofPseudomonas aeruginosa, 26Acinetobacter baumanniiisolates, and 11Stenotrophomonas maltophiliaisolates. In total, 15 antimicrobials were evaluated, with 11 forP. aeruginosa, 14 forA. baumannii, and 2 forS. maltophilia. Categorical agreement (CA) was assessed using both Vitek 2 breakpoints and 2016 CLSI M100S 26th edition breakpoints. The essential agreement values forP. aeruginosa,A. baumannii, andS. maltophiliawere 99.5%, 99.2%, and 100%, respectively. The CA values forP. aeruginosa,A. baumannii, andS. maltophiliawere 94.1%, 92.7%, and 95.5%, respectively, by the Vitek 2 breakpoints, and 93.4%, 92.3%, and 95.5%, respectively, by the CLSI breakpoints. Overall, the Vitek 2 performance was comparable to that of BMD using both Vitek 2 breakpoints and 2016 CLSI M100S 26th edition breakpoints. Improved performance was noted for the reformulated piperacillin-tazobactam and imipenem found on the AST-GN69 card, with no very major or major errors noted when using the CLSI breakpoints.

scholarly journals Specific and rapid reverse assaying protocol for detection and antimicrobial susceptibility testing of Pseudomonas aeruginosa based on dual molecular recognition

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yong He ◽  
Hang Zhao ◽  
Yuanwen Liu ◽  
He Zhou
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antimicrobial Susceptibility ◽  
Magnetic Particles ◽  
Susceptibility Testing ◽  
Fluorescein Isothiocyanate ◽  
High Specificity ◽  
Antimicrobial Susceptibility Testing ◽  
Tail Fiber ◽  
Isolation And Identification ◽  
Magainin Ii

AbstractThe worldwide emergence and spread of antimicrobial resistance is accelerated by irrational administration and use of empiric antibiotics. A key point to the crisis is a lack of rapid diagnostic protocols for antimicrobial susceptibility testing (AST), which is crucial for a timely and rational antibiotic prescription. Here, a recombinant bacteriophage tail fiber protein (TFP) was functionalized on magnetic particles to specifically capture Pseudomonas aeruginosa, while fluorescein isothiocyanate-labeled-magainin II was utilized as the indicator. For solving the magnetic particles’ blocking effects, a reverse assaying protocol based on TFP recognition was developed to investigate the feasibility of detection and AST of P. aeruginosa. P. aeruginosa can be rapidly, sensitively and specifically detected within 1.5 h with a linear range of 1.0 × 102 to 1.0 × 106 colony forming units (CFU)⋅mL−1 and a detection limit of 3.3 × 10 CFU⋅mL−1. Subsequently, AST results, which were consistent with broth dilution results, can be obtained within 3.5 h. Due to the high specificity of the TFP, AST can actually be conducted without the need for bacterial isolation and identification. Based on the proof-of-principle work, the detection and AST of other pathogens can be extended by expressing the TFPs of their bacteriophages.

scholarly journals The Slow March toward Rapid Phenotypic Antimicrobial Susceptibility Testing: Are We There Yet?

2018 ◽  
Vol 56 (4) ◽  
pp. e01999-17 ◽  
Author(s):  
Christopher D. Doern
Keyword(s):  
Antimicrobial Susceptibility ◽  
Rapid Growth ◽  
Gold Standard ◽  
Historical Perspective ◽  
Susceptibility Testing ◽  
Turnaround Time ◽  
Antimicrobial Susceptibility Testing ◽  
Standard Methods ◽  
Phenotypic Susceptibility ◽  
Made In

ABSTRACT Antimicrobial susceptibility testing (AST) provides critical information for the management of patients with infections. The gold standard methods for assessing organism susceptibility are still based on growth and require incubation over relatively long periods of time. Until now, little progress has been made in developing rapid, growth-based, phenotypic AST systems. This commentary puts the recently FDA-cleared Accelerate PhenoTest (P. Pancholi et al., J Clin Microbiol 56:e01329-17, 2018, https://doi.org/10.1128/JCM.01329-17) in context by providing a historical perspective on attempts to accelerate phenotypic susceptibility results. In addition, some promising new innovations that promise to shorten the turnaround time for phenotypic AST will be briefly reviewed.

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