scholarly journals On the Consequences of Poorly Defined Breakpoints for Rifampin Susceptibility Testing of Mycobacterium tuberculosis Complex

2021 ◽  
Vol 59 (4) ◽  
Author(s):  
Claudio U. Köser ◽  
Sophia B. Georghiou ◽  
Thomas Schön ◽  
Max Salfinger
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
World Health ◽  
The European Union ◽  
Reference Standard ◽  
Resistance Mutations ◽  
Content Type ◽  
Indicator Tube

ABSTRACT In a recent report of a systematic review of critical concentrations (CCs), the World Health Organization (WHO) lowered the rifampin (RIF) CC for antimicrobial susceptibility testing (AST) of the Mycobacterium tuberculosis complex using Middlebrook 7H10 medium and the Bactec Mycobacterial Growth Indicator Tube (MGIT) 960 system from 1 to 0.5 μg/ml. The previous RIF CC for 7H10 had been in use for over half a century. Because it had served as the de facto reference standard, it contributed to the endorsement of inappropriately high CCs for other AST methods, including the U.S. Food and Drug Administration (FDA)-approved MGIT system. Moreover, this resulted in confusion about the interpretation of seven borderline resistance mutations in rpoB (i.e., L430P, D435Y, H445L, H445N, H445S, L452P, and I491F). In this issue of the Journal of Clinical Microbiology, Shea et al. (J Clin Microbiol 59:e01885-20, 2021, https://doi.org/10.1128/JCM.01885-20) provide evidence that the CC endorsed by the Clinical and Laboratory Standards Institute for the Sensititre MYCOTB system, which is not FDA approved but is CE-IVD marked in the European Union, is likely also too high. These findings underscore the importance of calibrating AST methods against a rigorously defined reference standard, as recently proposed by the European Committee on Antimicrobial Susceptibility Testing, as well as the value of routine next-generation sequencing for investigating discordant AST results.

scholarly journals Analysis of Whole-Genome Sequences for the Prediction of Penicillin Resistance and β-Lactamase Activity inBacillus anthracis

mSystems ◽  
2018 ◽  
Vol 3 (6) ◽  
Author(s):  
A. S. Gargis ◽  
H. P. McLaughlin ◽  
A. B. Conley ◽  
C. Lascols ◽  
P. A. Michel ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Antimicrobial Susceptibility ◽  
Sigma Factor ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Penicillin Resistance ◽  
Whole Genome ◽  
Activity Levels ◽  
Content Type

ABSTRACTPenicillin (PEN) is a low-cost option for anthrax treatment, but naturally occurring resistance has been reported. β-Lactamase expression (bla1,bla2) inBacillus anthracisis regulated by a sigma factor (SigP) and its cognate anti-sigma factor (RsiP). Mutations leading to truncation of RsiP were previously described as a basis for PEN resistance. Here, we analyze whole-genome sequencing (WGS) data and compare the chromosomalsigP-bla1regions from 374B. anthracisstrains to determine the frequency of mutations, identify mutations associated with PEN resistance, and evaluate the usefulness of WGS for predicting PEN resistance. Few (3.5%) strains contained at least 1 of 11 different mutations insigP,rsiP, orbla1.Nine of these mutations have not been previously associated with PEN resistance. Four strains showed PEN resistance (PEN-R) by conventional broth microdilution, including 1 strain with a novel frameshift inrsiP. One strain that carries the samersiPframeshift mutation as that found previously in a PEN-R strain showed a PEN-susceptible (PEN-S) phenotype and exhibited decreasedbla1andbla2transcription. An unexpectedly small colony size, a reduced growth rate, and undetectable β-lactamase activity levels (culture supernatant and cell lysate) were observed in this PEN-S strain. Sequence analysis revealed mutations in genes associated with growth defects that may contribute to this phenotype. WhileB. anthracisrsiPmutations cannot be exclusively used to predict resistance, four of the five strains withrsiPmutations were PEN-R. Therefore, theB. anthracissigP-bla1region is a useful locus for WGS-based PEN resistance prediction, but phenotypic testing remains essential.IMPORTANCEDetermination of antimicrobial susceptibility ofB. anthracisis essential for the appropriate distribution of antimicrobial agents for postexposure prophylaxis (PEP) and treatment of anthrax. Analysis of WGS data allows for the rapid detection of mutations in antimicrobial resistance (AMR) genes in an isolate, but the presence of a mutation in an AMR gene does not always accurately predict resistance. As mutations in the anti-sigma factor RsiP have been previously associated with high-level penicillin resistance in a limited number of strains, we investigated WGS assemblies from 374 strains to determine the frequency of mutations and performed functional antimicrobial susceptibility testing. Of the five strains that contained mutations inrsiP, only four were PEN-R by functional antimicrobial susceptibility testing. We conclude that while sequence analysis of this region is useful for AMR prediction inB. anthracis, genetic analysis should not be used exclusively and phenotypic susceptibility testing remains essential.

scholarly journals Tailoring Antimicrobial Susceptibility Testing to Individual Species of Coagulase-Negative Staphylococci: Next Up, Staphylococcus epidermidis

2019 ◽  
Vol 57 (12) ◽  
Author(s):  
C. Paul Morris ◽  
Patricia J. Simner
Keyword(s):  
Antimicrobial Susceptibility ◽  
Staphylococcus Epidermidis ◽  
Susceptibility Testing ◽  
Methicillin Resistance ◽  
Antimicrobial Susceptibility Testing ◽  
Individual Species ◽  
Beta Lactam ◽  
Coagulase Negative Staphylococci ◽  
Content Type ◽  
Species Specific

ABSTRACT Accurate detection of methicillin resistance among staphylococci is vital for patient care. Methicillin resistance is most commonly mediated by acquisition of the mecA gene, which encodes an altered penicillin binding protein, PBP2a. Application of phenotypic methods to detect mecA-mediated beta-lactam resistance in staphylococci is becoming more complex as species-specific differences are identified among coagulase-negative staphylococci (CoNS). Previously, interpretative criteria and antimicrobial susceptibility testing (AST) methods specific to the CoNS group were used to evaluate Staphylococcus epidermidis. A manuscript by S. N. Naccache, K. Callan, C.-A. D. Burnham, M. A. Wallace, et al. (J Clin Microbiol 57:e00961-19, 2019, https://doi.org/10.1128/JCM.00961-19) details experiments revealing that S. epidermidis, the most common clinically isolated CoNS, requires tailored use of previously described methods and interpretive criteria to reliably identify the presence of mecA-mediated methicillin resistance.

scholarly journals Evaluation of the Accelerate Pheno System for Fast Identification and Antimicrobial Susceptibility Testing from Positive Blood Cultures in Bloodstream Infections Caused by Gram-Negative Pathogens

2017 ◽  
Vol 55 (7) ◽  
pp. 2116-2126 ◽  
Author(s):  
Matthias Marschal ◽  
Johanna Bachmaier ◽  
Ingo Autenrieth ◽  
Philipp Oberhettinger ◽  
Matthias Willmann ◽  
...  
Keyword(s):  
Blood Culture ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Bloodstream Infections ◽  
Test System ◽  
Blood Cultures ◽  
Antimicrobial Susceptibility Testing ◽  
Diagnostic Tools ◽  
Gram Negative ◽  
Content Type

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.

scholarly journals Rapid Antimicrobial Susceptibility Testing Using Forward Laser Light Scatter Technology

2016 ◽  
Vol 54 (11) ◽  
pp. 2701-2706 ◽  
Author(s):  
Randall T. Hayden ◽  
Lani K. Clinton ◽  
Carolyn Hewitt ◽  
Terri Koyamatsu ◽  
Yilun Sun ◽  
...  
Keyword(s):  
Antimicrobial Susceptibility ◽  
Clinical Decision Making ◽  
Susceptibility Testing ◽  
Laser Light ◽  
Accurate Determination ◽  
Antimicrobial Susceptibility Testing ◽  
Limiting Factor ◽  
Light Scatter ◽  
Content Type

The delayed reporting of antimicrobial susceptibility testing remains a limiting factor in clinical decision-making in the treatment of bacterial infection. This study evaluates the use of forward laser light scatter (FLLS) to measure bacterial growth for the early determination of antimicrobial susceptibility. Three isolates each (two clinical isolates and one reference strain) ofStaphylococcus aureus,Escherichia coli, andPseudomonas aeruginosawere tested in triplicate using two commercial antimicrobial testing systems, the Vitek2 and the MicroScan MIC panel, to challenge the BacterioScan FLLS. The BacterioScan FLLS showed a high degree of categorical concordance with the commercial methods. Pairwise comparison with each commercial system serving as a reference standard showed 88.9% agreement with MicroScan (two minor errors) and 72.2% agreement with Vitek (five minor errors). FLLS using the BacterioScan system shows promise as a novel method for the rapid and accurate determination of antimicrobial susceptibility.

scholarly journals Wild-Type and Non-Wild-Type Mycobacterium tuberculosis MIC Distributions for the Novel Fluoroquinolone Antofloxacin Compared with Those for Ofloxacin, Levofloxacin, and Moxifloxacin

2016 ◽  
Vol 60 (9) ◽  
pp. 5232-5237 ◽  
Author(s):  
Xia Yu ◽  
Guirong Wang ◽  
Suting Chen ◽  
Guomei Wei ◽  
Yuanyuan Shang ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Critical Concentration ◽  
Bacterial Species ◽  
Drug Susceptibility Testing ◽  
World Health ◽  
Wild Type ◽  
Resistance Mutations ◽  
Content Type ◽  
Lowenstein Jensen

ABSTRACTAntofloxacin (AFX) is a novel fluoroquinolone that has been approved in China for the treatment of infections caused by a variety of bacterial species. We investigated whether it could be repurposed for the treatment of tuberculosis by studying itsin vitroactivity. We determined the wild-type and non-wild-type MIC ranges for AFX as well as ofloxacin (OFX), levofloxacin (LFX), and moxifloxacin (MFX), using the microplate alamarBlue assay, of 126 clinicalMycobacterium tuberculosisstrains from Beijing, China, of which 48 were OFX resistant on the basis of drug susceptibility testing on Löwenstein-Jensen medium. The MIC distributions were correlated with mutations in the quinolone resistance-determining regions ofgyrA(Rv0006) andgyrB(Rv0005). Pharmacokinetic/pharmacodynamic (PK/PD) data for AFX were retrieved from the literature. AFX showed lower MIC levels than OFX but higher MIC levels than LFX and MFX on the basis of the tentative epidemiological cutoff values (ECOFFs) determined in this study. All strains with non-wild-type MICs for AFX harbored known resistance mutations that also resulted in non-wild-type MICs for LFX and MFX. Moreover, our data suggested that the current critical concentration of OFX for Löwenstein-Jensen medium that was recently revised by the World Health Organization might be too high, resulting in the misclassification of phenotypically non-wild-type strains with known resistance mutations as wild type. On the basis of our exploratory PK/PD calculations, the current dose of AFX is unlikely to be optimal for the treatment of tuberculosis, but higher doses could be effective.

scholarly journals What's in a Name? The Impact of Accurate Staphylococcus pseudintermedius Identification on Appropriate Antimicrobial Susceptibility Testing

2016 ◽  
Vol 54 (3) ◽  
pp. 516-517 ◽  
Author(s):  
Brandi M. Limbago
Keyword(s):  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Methicillin Resistance ◽  
Antimicrobial Susceptibility Testing ◽  
Robust Identification ◽  
Content Type ◽  
Staphylococcus Intermedius ◽  
Article Type ◽  
The Impact ◽  
Coagulase Positive Staphylococci

Bacteria in theStaphylococcus intermediusgroup, includingStaphylococcuspseudintermedius, often encodemecA-mediated methicillin resistance. Reliable detection of this phenotype for proper treatment and infection control decisions requires that these coagulase-positive staphylococci are accurately identified and specifically that they are not misidentified asS. aureus. As correct species level bacterial identification becomes more commonplace in clinical laboratories, one can expect to see changes in guidance for antimicrobial susceptibility testing and interpretation. The study by Wu et al. in this issue (M. T. Wu, C.-A. D. Burnham, L. F. Westblade, J. Dien Bard, S. D. Lawhon, M. A. Wallace, T. Stanley, E. Burd, J. Hindler, R. M. Humphries, J Clin Microbiol 54:535–542, 2016,http://dx.doi.org/10.1128/JCM.02864-15) highlights the impact of robust identification ofS. intermediusgroup organisms on the selection of appropriate antimicrobial susceptibility testing methods and interpretation.

scholarly journals Antimicrobial resistance surveillance in the South African private sector report for 2016

2018 ◽  
Vol 33 (4) ◽  
pp. 114-117
Author(s):  
Olga Perovic ◽  
Husna Ismail ◽  
Erika Van Schalkwyk ◽  
Warren Lowman ◽  
Elizabeth Prentice ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Private Sector ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Action Plan ◽  
Treatment Resistance ◽  
Antimicrobial Susceptibility Testing ◽  
World Health ◽  
Global Action ◽  
High Level

Aim: The relevance of surveillance for antimicrobial resistance is increasingly recognised in the light of a global action plan to combat resistance. This report presents antimicrobial susceptibility testing on ESKAPE pathogens from private sector laboratories in South Africa for 2016.Methods: Antimicrobial susceptibility testing (AST) performed on ESKAPE organisms (Enterococcus faecium, Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter cloacae and Escherichia coli) isolated from blood cultures at four private pathology laboratories in 2016 were analysed. Analysis and reporting of data were done via a uniform platform created by the NICD for national AST data.Results: AST were reported on 9 029 ESKAPE organisms including 58% Enterobacteriaceae, 28% Gram-positive bacteria and 14% Gram-negative bacteria and drug-bug combination was performed following the Global Antimicrobial Surveillance System (GLASS) guidelines by the World Health Organization.Conclusions: The most important resistance to address is a high level of ESBL in Enterobacteriaceae, which necessitates the use of carbapenems for treatment. Resistance to carbapenems is recorded in this report but not confirmation of genes by genotypic methods. During this period, no increase in vancomycin-resistant Enterococci was observed.

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.

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