Accurate Antimicrobial Susceptibility Testing Research Articles

Sliding motility of Bacillus cereus mediates vancomycin pseudo-resistance during antimicrobial susceptibility testing.

The glycopeptide vancomycin is the antimicrobial agent-of-choice for the treatment of severe non-gastrointestinal infections with members of Bacillus cereus sensu lato (s.l.). Recently, sporadic detection of vancomycin-resistant phenotypes emerged, mostly for agar diffusion testing such as the disc diffusion method or gradient test (e.g. Etest®) method. In this work, we were able to disprove a preliminarily assumed high resistance to vancomycin in an isolate of B. cereus s.l. using broth microdilution and agar dilution. Microscopic imaging during vancomycin susceptibility testing showed spreading towards the inhibition zone, which strongly suggested sliding motility. Furthermore, transcriptomic analysis using RNA-Seq on the nanopore platform revealed several key genes of biofilm formation (e.g. calY, tasA, krsEABC) to be up-regulated in pseudo-resistant cells, substantiating that bacterial sliding is responsible for the observed mobility. Down-regulation of virulence (e.g. hblABCD, nheABC, plcR) and flagellar genes compared with swarming cells also confirmed the non-swarming phenotype of the pseudo-resistant isolate. The results highlight an insufficiency of agar diffusion testing for vancomycin susceptibility in the B. cereus group, and reference methods like broth microdilution are strongly recommended. As currently no guideline mentions interfering phenotypes in antimicrobial susceptibility testing of B. cereus s.l., this knowledge is essential to obtain reliable results on vancomycin susceptibility. In addition, this is the first report of sliding motility undermining accurate antimicrobial susceptibility testing in B. cereus s.l. and may serve as a basis for future studies on bacterial motility in susceptibility testing and its potential impact on treatment efficacy.

Rapid determination of antimicrobial susceptibility of Gram-negative bacteria from clinical blood cultures using a scattered light-integrated collection device.

Background. A bloodstream infection (BSI) presents a complex and serious health problem, a problem that is being exacerbated by increasing antimicrobial resistance (AMR).Gap Statement. The current turnaround times (TATs) for most antimicrobial susceptibility testing (AST) methods offer results retrospective of treatment decisions, and this limits the impact AST can have on antibiotic prescribing and patient care. Progress must be made towards rapid BSI diagnosis and AST to improve antimicrobial stewardship and reduce preventable deaths from BSIs. To support the successful implementation of rapid AST (rAST) in hospital settings, a rAST method that is affordable, is sustainable and offers comprehensive AMR detection is needed.Aim. To evaluate a scattered light-integrated collection (SLIC) device against standard of care (SOC) to determine whether SLIC could accelerate the current TATs with actionable, accurate rAST results for Gram-negative BSIs.Methods. Positive blood cultures from a tertiary referral hospital were studied prospectively. Flagged positive Gram-negative blood cultures were confirmed by Gram staining and analysed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, Vitek 2, disc diffusion (ceftriaxone susceptibility only) and an SLIC device. Susceptibility to a panel of five antibiotics, as defined by European Committee on Antimicrobial Susceptibility Testing breakpoints, was examined using SLIC.Results. A total of 505 bacterial-antimicrobial combinations were analysed. A categorical agreement of 95.5 % (482/505) was achieved between SLIC and SOC. The 23 discrepancies that occurred were further investigated by the broth microdilution method, with 10 AST results in agreement with SLIC and 13 in agreement with SOC. The mean time for AST was 10.53±0.46 h and 1.94±0.02 h for Vitek 2 and SLIC, respectively. SLIC saved 23.96±1.47 h from positive blood culture to AST result.Conclusion. SLIC has the capacity to provide accurate AST 1 day earlier from flagged positive blood cultures than SOC. This significant time saving could accelerate time to optimal antimicrobial therapy, improving antimicrobial stewardship and management of BSIs.

The Potential Clinical Applications of a Microfluidic Lab-on-a-Chip for the Identification and Antibiotic Susceptibility Testing of Enterococcus faecalis-Associated Endodontic Infections: A Systematic Review.

This systematic review evaluated the potential clinical use of microfluidic lab-on-a-chip (LOC) technology in the identification and antibiotic susceptibility testing of E. faecalis in endodontic infections. The search methodology employed in this review adhered to the PRISMA guidelines. Multiple scientific databases, including PubMed/MEDLINE, SCOPUS, and SCIELO, were utilized, along with exploration of grey literature sources. Up to September 2023, these resources were searched using specific keywords and MeSH terms. An initial comprehensive search yielded 202 articles. Ultimately, this systematic review incorporated 12 studies. Out of these, seven aimed to identify E. faecalis, while the remaining five evaluated its susceptibility to different antibiotics. All studies observed that the newly developed microfluidic chip significantly reduces detection time compared to traditional methods. This enhanced speed is accompanied by a high degree of accuracy, efficiency, and sensitivity. Most research findings indicated that the entire process took anywhere from less than an hour to five hours. It is important to note that this approach bypasses the need for minimum inhibitory concentration measurements, as it does not rely on traditional methodologies. Microfluidic devices enable the rapid identification and accurate antimicrobial susceptibility testing of E. faecalis, which are crucial for timely diagnosis and treatment in endodontic infections.

Updating breakpoints in the United States: a summary from the ASM Clinical Microbiology Open 2022.

Accurate antimicrobial susceptibility testing (AST) and reporting are essential for guiding appropriate therapy for patients and direction for public health prevention and control actions. A critical feature of AST reporting is the interpretation of AST results using clinical breakpoints for reporting as susceptible, susceptible-dose dependent, intermediate, or resistant. Breakpoints are subject to continuous adjustment and updating to best reflect current clinical data. These breakpoint changes can benefit patients and public health only if adopted in a timely manner. A recent survey identified that up to 70% of College of American Pathologists (CAP)-accredited U.S. laboratories and 45% of CAP-accredited laboratories outside the U.S. use various obsolete clinical breakpoints to interpret AST results to guide patient care. The reason for the ongoing use of obsolete breakpoints is multifactorial, including barriers encountered by laboratories, commercial AST device manufacturers, standards development organizations, and regulatory bodies alike. To begin to address this important patient safety issue, CAP implemented checklist requirements for CAP-accredited laboratories to ensure up-to-date clinical breakpoint use. Furthermore, the topic was discussed at the June 2022 American Society for Microbiology Clinical Microbiology Open (CMO) with various stakeholders to identify potential solutions. This minireview summarizes the breakpoint setting process in the U.S. and highlights solutions to close the gap between breakpoint revisions and implementation in clinical and public health laboratories. Solutions discussed include clarification of data requirements and minimum inhibitory concentration only reporting for regulatory clearance of AST devices, clinical data generation to close breakpoints gaps, advocacy, education, and greater dialogue between stakeholders.

Rapid and Accurate Antimicrobial Susceptibility Testing Using Label-Free Electrical Impedance-Based Microfluidic Platform.

Antimicrobial resistance has become a serious threat to the global public health. Accurate and rapid antimicrobial susceptibility testing (AST) allows evidence-based prescribing of antibiotics to improve patient care and clinical outcomes. Current culture-based AST assays are inherently limited by the doubling time of bacterial reproduction, which require at least 24 h to have a decisive result. Herein, a label-free electrical impedance-based microfluidic platform designed to expedite and streamline AST procedure for clinical practice is presented. Following a 30-min exposure of bacterial samples to antibiotics, the presented high-throughput, single-bacterium level impedance characterization platform enables a rapid 2-min AST assay. The platform facilitates accurate analysis of individual bacterial viability, as indicated by changes in electrical characteristics, thereby enabling the determination of antimicrobial resistance. Moreover, the potential clinical applicability of this platform is demonstrated by testing different E. coli strains against five antibiotics, yielding 100% categorical agreements compared to standard culture methods.

A-274 Validation of Cefepime-Taniborbactam MIC Antimicrobial Susceptibility Test for MicroScan Dried Gram-Negative MIC Panels from a Multicenter Assessment of Enterobacterales and Pseudomonas aeruginosa

Abstract Background Cefepime-taniborbactam is an investigational agent with activity against carbapenem and multidrug-resistant Enterobacterales and Pseudomonas aeruginosa.1 Given the need for accurate antimicrobial susceptibility testing results to support patient care decisions, a multicenter study was conducted with Enterobacterales and Pseudomonas aeruginosa clinical isolates on investigational MicroScan Dried Gram-negative MIC (MSDGN) panels with cefepime-taniborbactam.2 Materials/Methods MSDGN panels were evaluated at three clinical sites by comparing MIC values obtained using MSDGN panels to MICs utilizing CLSI broth microdilution reference panels. The study included 490/491 Enterobacterales and 65 Pseudomonas aeruginosa clinical isolates tested using Prompt® and turbidity methods of inoculation during the combined efficacy and challenge phases. MSDGN panels were incubated at 35 ± 1°C and read on the WalkAway System, the autoSCAN-4 instrument, and visually at 16–20 h. Frozen reference panels were prepared according to CLSI/ISO methodology, incubated for 16–20 h and read visually. Results Essential agreement was calculated compared to MIC results from frozen reference panels for isolates tested in efficacy and challenge and found in the following table. Conclusion Cefepime-taniborbactam MIC results for Enterobacterales and Pseudomonas aeruginosa obtained with the MSDGN panel correlate well with MICs obtained using frozen reference panels in this multicenter study.Pending approval by the US Food and Drug Administration; not available for in vitro diagnostic use in the US. Not for Distribution in the US.Pending submission and clearance by the US Food and Drug Administration; not yet available for in vitro diagnostic use in the US.For Investigational Use Only. Performance characteristics of this product have not been established.Read methodEnterobacterales Essential agreement (EA) % (n/N)Pseudomonas aeruginosaEssential agreement (EA) % (n/N)PromptTurbidityPromptTurbidityWalkAway90.0 (441/490)99.2 (487/491)100.0 (65/65)100.0 (65/65)autoSCAN-494.3 (462/490)99.0 (486/491)98.5 (64/65)98.5 (64/65)Visually92.7 (454/490)99.2 (487/491)100.0 (65/65)100.0 (65/65)Abbreviations: Prompt, Prompt inoculation method, Turbidity, Turbidity inoculation method.

Differences in antimicrobial susceptibility testing complicating management of IMP carbapenemase-producing Enterobacterales infection

IMP-type carbapenemases are rarely detected in Europe and limited information is available to guide the treatment of infections caused by carbapenemase-producing Enterobacterales (CPE) producing these carbapenemases. Accurate antimicrobial susceptibility testing (AST) results are essential for optimal antibiotic management. Here we report discrepancies in AST of IMP-producing Enterobacterales (IMP-CPE) complicating the management of severe sepsis. Antimicrobial susceptibilities were analysed by in-house VITEK® 2, Etest and broth microdilution (BMD). Carbapenemase-encoding genes were detected by PCR. Whole-genome sequencing (WGS) was performed using an Illumina MiSeq platform. Minimum inhibitory concentrations (MICs) determined by VITEK® 2 for Enterobacter hormaechei and Klebsiella oxytoca blood culture isolates were ≥16 mg/L for meropenem and ≤0.5 mg/L for ertapenem. In contrast, Etest analysis and BMD returned MICs of 2 mg/L and 1 mg/L, respectively. Both isolates tested positive for IMP carbapenemase-encoding genes by PCR. WGS revealed that both isolates carried the same blaIMP-4 gene. Based on VITEK® 2 susceptibilities, initial treatment was with tigecycline and amikacin. After subsequent deterioration, the patient was successfully treated with ertapenem and amikacin. This case highlights that automated AST by VITEK® 2 can over-report meropenem resistance for IMP carbapenemase-producers compared with Etest and BMD. Clinicians need to be cautious deciding against carbapenem treatment based on VITEK® 2 susceptibility testing results for IMP-positive Enterobacterales. Tigecycline was inferior to carbapenem treatment for pyelonephritis caused by isolates expressing IMP carbapenemases, however specific evidence guiding the treatment of these infections is lacking.

Interference Disturbance Analysis Enables Single-Cell Level Growth and Mobility Characterization for Rapid Antimicrobial Susceptibility Testing.

To support the emerging battle against antimicrobial resistance (AMR), detection methods that allow fast and accurate antimicrobial susceptibility testing (AST) are urgently needed. The early identification and application of an appropriate antibiotic treatment leads to lower mortality rates and substantial cost savings and prevents the development of resistant pathogens. In this work, we present a diffraction-based method, which is capable of quantitative bacterial growth, mobility, and susceptibility measurements. The method is based on the temporal analysis of the intensity of a light diffraction peak, which arises due to interference at a periodic pattern of gold nanostructures. The presence of bacteria disturbs the constructive interference, leading to an intensity decrease and thus allows the monitoring of bacterial growth in very low volumes. We demonstrate the direct correlation of the decrease in diffraction peak intensity with bacterial cell number starting from single cells and show the capability for rapid high-throughput AST measurements by determining the minimum inhibitory concentration for three different antimicrobials in less than 2-3 h as well as the susceptibility in less than 30-40 min. Furthermore, bacterial mobility is obtained from short-term fluctuations of the diffraction peak intensity and is shown to decrease by a factor of 3 during bacterial attachment to a surface. This multiparameter detection method allows for rapid AST of planktonic and of biofilm-forming bacterial strains in low volumes and in real-time without the need of high initial cell numbers.

Performance of microbial identification by MALDI-TOF MS and susceptibility testing by VITEK 2 from positive blood cultures after minimal incubation on solid media.

Bloodstream infections (BSIs) are a leading cause of patient morbidity and mortality. Rapid identification of organisms from BSIs is critical for initiating targeted antimicrobial therapy. Although many methods exist for rapid identification, they do not provide detailed or definitive susceptibility information. We assessed the utility of both the VITEK MS and Bruker Biotyper MALDI-TOF mass spectrometers to identify organisms from a positive blood culture bottle after only 4 h of growth on solid media compared to identification from overnight growth using the VITEK MS. Additionally, we determined whether this limited growth could yield accurate antimicrobial susceptibility testing (AST) results compared to overnight growth using the VITEK 2 AST system. Overall, identifications using the VITEK MS and Biotyper had agreements of 127/150 (84%) and 133/150 (88%), respectively. For rapid AST, the overall categorical agreement was 1010/1017 (99.3%), where Gram-negative bacteria had concordant results for 743/750 (99.1%) organism-drug combinations and Gram-positive bacteria had concordant results for 265/267 (99.3%). Gram-negative bacteria had 4, 2, and 1 minor, major, and very major discrepancies, respectively, while Gram-positive bacteria had no minor errors, one major, and one very major discrepancy. In conclusion, organisms grown for only 4 h on solid media were accurately identified by MALDI-TOF MS and have concordant phenotypic AST profiles. This method can also be implemented using common commercial instruments, providing a way to improve upon identification and gain detailed susceptibility information without significant additional laboratory costs.

A new rapid method for direct antimicrobial susceptibility testing of bacteria from positive blood cultures.

BackgroundRapid identification and antimicrobial susceptibility testing (AST) of the causative agent(s) of bloodstream infections can lead to prompt appropriate antimicrobial therapy. To shorten species identification, in this study bacteria were recovered from monomicrobial blood cultures by serum separator tubes and spotted onto the target plate for direct MALDI-TOF MS identification. Proper antibiotics were selected for direct AST based on species identification. In order to obtain rapid AST results, bacteria were recovered from positive blood cultures by two different protocols: by serum separator tubes (further referred to as PR1), or after a short-term subculture in liquid medium (further referred to as PR2). The results were compared with those obtained by the method currently used in our laboratory consisting in identification by MALDI-TOF and AST by Vitek 2 or Sensititre on isolated colonies.ResultsThe direct MALDI-TOF method concordantly identified with the current method 97.5 % of the Gram-negative bacteria and 96.1 % of the Gram-positive cocci contained in monomicrobial blood cultures. The direct AST by PR1 and PR2 for all isolate/antimicrobial agent combinations was concordant/correct with the current method for 87.8 and 90.5 % of Gram-negative bacteria and for 93.1 and 93.8 % of Gram-positive cocci, respectively. In particular, 100 % categorical agreement was found with levofloxacin for Enterobacteriaceae by both PR1 and PR2, and 99.0 and 100 % categorical agreement was observed with linezolid for Gram-positive cocci by PR1 and PR2, respectively. There was no significant difference in accuracy between PR1 and PR2 for Gram-negative bacteria and Gram-positive cocci.ConclusionsThis newly described method seems promising for providing accurate AST results. Most importantly, these results would be available in a few hours from blood culture positivity, which would help clinicians to promptly confirm or streamline an effective antibiotic therapy in patients with bloodstream infections.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-016-0805-5) contains supplementary material, which is available to authorized users.

Identification of Low-Level Vancomycin Resistance in Staphylococcus aureus in the Era of Informatics.

Vancomycin-intermediateStaphylococcus aureus(VISA) and heteroresistant VISA (hVISA) are pathogens for which accurate antimicrobial susceptibility testing (AST) would rule out standard treatment with vancomycin. Unfortunately, AST for vancomycin is relatively slow and standard methods are unable to reliably detect VISA and hVISA. An article in this issue (C. A. Mather, B. J. Werth, S. Sivagnanam, D. J. SenGupta, and S. M. Butler-Wu, J Clin Microbiol 54:883-890, 2016, doi:http://dx.doi.org/10.1128/JCM.02428-15) describes a rapid whole-cell matrix-assisted laser desorption ionization-time of flight proxy susceptibility method that highlights current innovations and challenges with rapid AST, VISA/hVISA identification, and clinical bioinformatics.

Bacterial Cytological Profiling (BCP) as a Rapid and Accurate Antimicrobial Susceptibility Testing Method for Staphylococcus aureus

Successful treatment of bacterial infections requires the timely administration of appropriate antimicrobial therapy. The failure to initiate the correct therapy in a timely fashion results in poor clinical outcomes, longer hospital stays, and higher medical costs. Current approaches to antibiotic susceptibility testing of cultured pathogens have key limitations ranging from long run times to dependence on prior knowledge of genetic mechanisms of resistance. We have developed a rapid antimicrobial susceptibility assay for Staphylococcus aureus based on bacterial cytological profiling (BCP), which uses quantitative fluorescence microscopy to measure antibiotic induced changes in cellular architecture. BCP discriminated between methicillin-susceptible (MSSA) and -resistant (MRSA) clinical isolates of S. aureus (n=71) within 1–2h with 100% accuracy. Similarly, BCP correctly distinguished daptomycin susceptible (DS) from daptomycin non-susceptible (DNS) S. aureus strains (n=20) within 30min. Among MRSA isolates, BCP further identified two classes of strains that differ in their susceptibility to specific combinations of beta-lactam antibiotics. BCP provides a rapid and flexible alternative to gene-based susceptibility testing methods for S. aureus, and should be readily adaptable to different antibiotics and bacterial species as new mechanisms of resistance or multidrug-resistant pathogens evolve and appear in mainstream clinical practice.

International dissemination of Escherichia coli strains with discrepant behaviour in phenotypic antimicrobial susceptibility tests

Preventing the dissemination of antimicrobial resistance depends on appropriate antibiotic stewardship and accurate antimicrobial susceptibility testing (AST). We report the international dissemination of Escherichia coli strains, showing discrepancies between reference methods when phenotypically tested for susceptibility to piperacillin/tazobactam (TZP). We demonstrate that these related strains are predisposed to problematic TZP AST interpretations.

Using Rapid Diagnostic Tests to Optimize Antimicrobial Selection in Antimicrobial Stewardship Programs

New advances in rapid diagnostic tests provide several collaborative opportunities for stewardship teams, particularly in processes where delay in the initiation of appropriate therapy has significant consequences for patient outcomes. Rapid molecular tests enhance one of the key functions of microbiology laboratories, which is to produce accurate organism identification and timely antimicrobial susceptibility testing data. These data are used to guide the choice of antimicrobial agents for treating infections in individual patients, and the aggregate data (i.e., hospital's antibiogram) are used to guide empiric therapy for infections before the availability of the patient's culture results and antimicrobial susceptibility testing data. In addition, accurate antimicrobial susceptibility testing data are critical for the overall success of a health care institution's antimicrobial stewardship program. Tests that provide accurate organism identification and antimicrobial susceptibility not only benefit the individual patient but also increase the effectiveness of antimicrobial stewardship programs. This review focuses on the use of molecular diagnostic methods to rapidly identify Staphylococcus species, Enterococcus faecalis,Enterococcus faecium, Escherichia coli,Klebsiella pneumoniae,Pseudomonas aeruginosa,Clostridium difficile, and Candida species from clinical samples and summarizes studies that describe the application of rapid diagnostic tests in antimicrobial stewardship programs.

Hospital-acquired infections in the neonatal intensive care unit- Klebsiella pneumoniae

Klebsiella pneumoniae is medically the most important organism of the Klebsiella species. It is responsible for a significant proportion of hospital-acquired infections including septicemias, urinary tract infections, pneumonia, and soft tissue infections especially in the immunocompromised hosts such as the neonate. The hands of healthcare workers and the gastrointestinal tract of hospitalized infants serve as reservoirs for the transmission of the organism and are responsible for multiple hospital outbreaks. In recent years, there has been an increase in the incidence of outbreaks caused by multidrug resistance K. pneumoniae organisms or the extended spectrum beta-lactamase (ESBL)-producing K. pneumoniae. The problems associated with extended spectrum beta-lactamase-producing organisms include difficulties in accurate antimicrobial susceptibility testing, limited treatment options and increased morbidity and perhaps mortality. Hence, prevention through implementation of strict infection control guidelines, effective hand washing and judicious use of antimicrobials such as third generation cephalosporins is important to effectively reduce the morbidity associated with this infection.

Antimicrobial Susceptibility Testing -- How Accurate Are Our Laboratories?

As the prevalence of antibiotic resistance in pathogenic bacteria increases, so does the importance of accurate antimicrobial susceptibility testing by

Evaluation of the PASCO strep plus broth microdilution antimicrobial susceptibility panels for testing Streptococcus pneumoniae and other Streptococcal species.

Antimicrobial resistance continues to increase worldwide among isolates of Streptococcus pneumoniae and other species of streptococci. Increasing rates of penicillin resistance, particularly in viridans group streptococci, and resistance to multiple classes of antimicrobial agents, including beta-lactams, macrolides, and fluoroquinolones, in pneumococci have increased the importance of having accurate antimicrobial susceptibility testing results for guiding therapy. One commercial method of assessing resistance in streptococci is the PASCO Strep Plus panel. This broth microdilution-based method has recently been expanded to include a variety of newer antimicrobial agents. Therefore, we compared the results of the new PASCO Strep Plus panels for 26 antimicrobial agents against the results generated using the National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution reference method for 75 pneumococci and 68 other streptococcal isolates. Only 4 (0.2%) very major errors (all with pneumococci and each with a different antimicrobial agent) were observed. There were 5 (0.3%) major errors observed with pneumococci (each with a different antimicrobial agent), but only 1 major error with nonpneumococcal streptococci. All of the very major and major errors resolved on retesting. Of the 65 (3.9%) and 17 (1.6%) minor errors observed with pneumococci and other streptococci, respectively, all were within 1 dilution of the broth microdilution reference MIC result. Thus, the PASCO Strep Plus panel has comparable accuracy to the NCCLS broth microdilution reference method.

Multicenter laboratory validation of susceptibility testing of Mycobacterium tuberculosis against classical second-line and newer antimicrobial drugs by using the radiometric BACTEC 460 technique and the proportion method with solid media.

In a large multicenter study involving six major study sites in the United States, Canada, and Europe, the susceptibilities of 272 Mycobacterium tuberculosis strains to classical second-line antituberculosis (anti-TB) drugs (capreomycin, cycloserine, ethionamide, and kanamycin) and newer compounds (amikacin, clofazimine, ofloxacin, and rifabutin) were determined by the radiometric BACTEC 460 procedure and the conventional proportion method on Middlebrook 7H10 agar. Previously established critical concentrations for classical second-line anti-TB drugs were compared with several concentrations in liquid medium to establish equivalence. MICs of newer compounds determined in liquid medium were either the same or up to four times lower than those determined in agar medium. After establishing critical concentrations (breakpoints) in the extended testing of clinical isolates, we obtained an excellent overall correlation between the two systems, with no errors with amikacin, kanamycin, and ofloxacin and very few major or very major errors with the other drugs; however, for cycloserine, no breakpoint concentration could be recommended due to repeatedly inconsistent results by both methods. Based on these data we conclude that the BACTEC 460 procedure is a simple and rapid method requiring 4 to 8 days on average to generate accurate antimicrobial susceptibility testing (AST) results for eight anti-TB drugs other than those considered primary ones. These data not only fill a major gap of knowledge regarding the critical test concentrations of secondary anti-TB drugs but also provide a baseline for future evaluations of M. tuberculosis AST with the more recently developed, nonradiometric broth-based culture systems.