Rapid Antimicrobial Susceptibility Testing Research Articles

Clinical Pilot of Bacterial Transcriptional Profiling as a Combined Genotypic and Phenotypic Antimicrobial Susceptibility Test.

Antimicrobial resistance is a growing health threat, but standard methods for determining antibiotic susceptibility are slow and can delay optimal treatment, which is especially consequential in severe infections such as bacteremia. Novel approaches for rapid susceptibility profiling have emerged that characterize either bacterial response to antibiotics (phenotype) or detect specific resistance genes (genotype). GoPhAST-R is a novel assay, performed directly on positive blood cultures, that integrates rapid transcriptional response profiling with detection of key resistance gene transcripts, thereby providing simultaneous data on both phenotype and genotype. Here, we performed the first clinical pilot of GoPhAST-R on 42 positive blood cultures: 26 growing Escherichia coli, 15 growing Klebsiella pneumoniae, and 1 with both. An aliquot of each positive blood culture was exposed to 9 different antibiotics, lysed, then underwent rapid transcriptional profiling on the NanoString® platform; results were analyzed using an in-house susceptibility classification algorithm. GoPhAST-R achieved 95% overall agreement with standard antimicrobial susceptibility testing methods, with the highest agreement for beta-lactams (98%) and the lowest for fluoroquinolones (88%). Epidemic resistance genes including the extended spectrum beta-lactamase bla CTX-M-15 and the carbapenemase bla KPC were also detected within the population. This study demonstrates the clinical feasibility of using transcriptional response profiling for rapid resistance determination, although further validation with larger and more diverse bacterial populations will be essential in future work. GoPhAST-R represents a promising new approach for rapid and comprehensive antibiotic susceptibility testing in clinical settings.

Rapid Antimicrobial Susceptibility Testing Using the MicroScan System: Performance Evaluation of a 4-Hour Bacterial Cultivation From Positive Blood Cultures.

A reliable and above all, rapid antimicrobial susceptibility test (AST) is required for the diganostics of blood stream infections (BSI). In this study, resistance testing using DxM MicroScan WalkAway (MicroScan) from a 4-h subculture is compared with the standard overnight culture (18-24h). Randomly selected positive blood cultures (PBC, n = 102) with gram-negative bacteria were included in the study. PBC were sub-cultured onto appropriate agar plates and AST by MicroScan was performed after 4 h of incubation and repeated after incubation for 18-24 h as standard. In a total of 1909 drug-strain pairs, the 4-h subculture approach showed a very high essential agreement (EA) (98.6%) and categorical agreement (CA) (97.1%) compared with the standard. The incidence of minor error (mE), major error (ME), very major error (VME), and adjusted very major error (aVME) was 1.1%, 0.4%, 12.9%, and 5.3%, respectively. In summary, the use of 4-h subcultures for resistance testing with the MicroScan offers a very reliable and easy to realize time saving when testing positive blood cultures with gram-negative bacteria.

ARTIFICIAL INTELLIGENCE APPLICATIONS IN CLINICAL MICROBIOLOGY LABORATORY

Artificial intelligence is becoming an increasingly important component of clinical microbiology informatics. Researchers, microbiologists, laboratorians, and diagnosticians are interested in AI-based testing because these applications have the potential to improve the turnaround time, quality, and cost of a test. Artificial intelligence which has gained importance in the laboratory, is used to support decision-making, identification and antimicrobial susceptibility testing with various technologies, image analyses, and MALDI-TOF-MS in medical microbiology and in infectious disease testing. Treatment of infections requires rapid and accurate identification and antimicrobial susceptibility testing. Modern artificial intelligence (AI) and machine-learning (ML) methods can now complete tasks with performance characteristic comparable to those of expert human operators. As a result, many healthcare fields combine these technologies, including in vitro diagnostics and, more broadly laboratory medicine, incorporate these technologies. These technologies are rapidly being developed and disclosed, but by comparison, their application so far has been limited. We need to further establish best practices and improve our information system and communications infrastructure to promote the implementation of reliable and advanced machine learning-based technologies. İnvolvement of the clinical microbiology laboratory community is essential to ensure that laboratory data is adequately accessible and thoughtfully incorporated into robust, safe and clinically effective ML-supported clinical diagnoses. This has been made possible by advances in modern computing and the widespread digitalization of health information. In this review, it is aimed to give information about to AI and its subfield of ML application areas in clinical microbiology laboratory.

Can flow cytometric measurements of reactive oxygen species levels determine minimal inhibitory concentrations and antibiotic susceptibility testing for Acinetobacter baumannii?

Current antimicrobial susceptibility testing (AST) requires 16-24 hours, delaying initiation of appropriate antibiotics. Hence, there is a need for rapid AST. This study aims to develop and evaluate the feasibility of a rapid flow cytometric AST assay to determine minimum inhibitory concentration (MIC) for carbapenem-resistant Acinetobacter baumannii (CRAB). Antibiotic exposure causes increased intracellular reactive oxygen species (ROS) in bacteria. We hypothesized that ROS can be used as a marker to determine MIC. We assessed three CRAB clinical isolates across fifteen antibiotics at various concentrations in a customized 96-well microtiter plate. The antibiotics assessed include amikacin, beta-lactams (ampicillin/sulbactam, aztreonam, cefepime, ceftolozane/tazobactam, doripenem, imipenem, meropenem, and piperacillin/tazobactam), levofloxacin, polymyxin B, rifampicin, trimethoprim/sulfamethoxazole, and tetracyclines (tigecycline and minocycline). These clinical CRAB isolates were assessed for ROS after antibiotic treatment. Increased ROS levels indicated by increased RedoxSensorTM Green (RSG) fluorescence intensity was assessed using flow cytometry (FCM). MIC was set as the lowest antibiotic concentration that gives a ≥1.5-fold increase in mode RSG fluorescence intensity (MICRSG). Accuracy of MICRSG was determined by comparing against microtiter broth dilution method performed under CLSI guidelines. ROS was deemed accurate in determining the MICs for β-lactams (83.3% accuracy) and trimethoprim/sulfamethoxazole (100% accuracy). In contrast, ROS is less accurate in determining MICs for levofloxacin (33.3% accuracy), rifampicin (0% accuracy), amikacin (33.3% accuracy), and tetracyclines (33.3% accuracy). Collectively, this study described an FCM-AST assay to determine antibiotic susceptibility of CRAB isolates within 5 hours, reducing turnaround time up to 19 hours.

Rapid, antibiotic incubation-free determination of tuberculosis drug resistance using machine learning and Raman spectroscopy

Tuberculosis (TB) is the world's deadliest infectious disease, with over 1.5 million deaths and 10 million new cases reported anually. The causative organism Mycobacterium tuberculosis (Mtb) can take nearly 40 d to culture, a required step to determine the pathogen's antibiotic susceptibility. Both rapid identification and rapid antibiotic susceptibility testing of Mtb are essential for effective patient treatment and combating antimicrobial resistance. Here, we demonstrate a rapid, culture-free, and antibiotic incubation-free drug susceptibility test for TB using Raman spectroscopy and machine learning. We collect few-to-single-cell Raman spectra from over 25,000 cells of the Mtb complex strain Bacillus Calmette-Guérin (BCG) resistant to one of the four mainstay anti-TB drugs, isoniazid, rifampicin, moxifloxacin, and amikacin, as well as a pan-susceptible wildtype strain. By training a neural network on this data, we classify the antibiotic resistance profile of each strain, both on dried samples and on patient sputum samples. On dried samples, we achieve >98% resistant versus susceptible classification accuracy across all five BCG strains. In patient sputum samples, we achieve ~79% average classification accuracy. We develop a feature recognition algorithm in order to verify that our machine learning model is using biologically relevant spectral features to assess the resistance profiles of our mycobacterial strains. Finally, we demonstrate how this approach can be deployed in resource-limited settings by developing a low-cost, portable Raman microscope that costs <$5,000. We show how this instrument and our machine learning model enable combined microscopy and spectroscopy for accurate few-to-single-cell drug susceptibility testing of BCG.

Short-term culture for rapid identification by mass spectrometry and automated antimicrobial susceptibility testing from positive bottles

BackgroundEarly and appropriate antibiotic treatment improves the clinical outcome of patients with sepsis. There is an urgent need for rapid identification (ID) and antimicrobial susceptibility testing (AST) of bacteria that cause bloodstream infection (BSI). Rapid ID and AST can be achieved by short-term incubation on solid medium of positive blood cultures using MALDI-TOF mass spectrometry (MS) and the BD M50 system. The purpose of this study is to evaluate the performance of rapid method compared to traditional method.MethodsA total of 124 mono-microbial samples were collected. Positive blood culture samples were short-term incubated on blood agar plates and chocolate agar plates for 5 ∼ 7 h, and the rapid ID and AST were achieved through Zybio EXS2000 MS and BD M50 System, respectively.ResultsCompared with the traditional 24 h culture for ID, this rapid method can shorten the cultivation time to 5 ∼ 7 h. Accurate organism ID was achieved in 90.6% of Gram-positive bacteria (GP), 98.5% of Gram-negative bacteria (GN), and 100% of fungi. The AST resulted in the 98.5% essential agreement (EA) and 97.1% category agreements (CA) in NMIC-413, 99.4% EA and 98.9% CA in PMIC-92, 100% both EA and CA in SMIC-2. Besides, this method can be used for 67.2% (264/393) of culture bottles during routine work. The mean turn-around time (TAT) for obtaining final results by conventional method is approximately 72.6 ± 10.5 h, which is nearly 24 h longer than the rapid method.ConclusionsThe newly described method is expected to provide faster and reliable ID and AST results, making it an important tool for rapid management of blood cultures (BCs). In addition, this rapid method can be used to process most positive blood cultures, enabling patients to receive rapid and effective treatment.

Micro-flow cell washing technique combined with single-cell Raman spectroscopy for rapid and automatic antimicrobial susceptibility test of pathogen in urine

Facing the rapid spread of antimicrobial resistance, methods based on single-cell Raman spectroscopy have proven their advances in reducing the turn-around time (TAT) of antimicrobial susceptibility tests (AST). However, the Raman-based methods are still hindered by the prolonged centrifugal cell washing procedure, which may require complex labor operation and induce high mechanical stress, resulting in a pretreatment time of over 1 h as well as a high cell-loss probability. In this study, we developed a micro-flow cell washing device and corresponding Raman-compatible washing chips, which were able to automatically remove the impurities in the samples, retain the bacterial cell and perform Raman spectra acquisition in situ. Results of washing the 5- and 10-μm polymethyl methacrylate (PMMA) microspheres showed that the novel technique achieved a successful removal of 99 % impurity and an 80 % particle retention rate after 6 to 10 cycles of washing. The micro-flow cell washing technique could complete the pretreatment for urine samples in a 96-well plate within 10 min, only taking 15 % of the handling time required by centrifugation. The AST profiles of urine sample spiked with E. coli 25922, E. faecalis 29212, and S. aureus 29213 obtained by the proposed Raman-based approach were found to be 100 % consistent with the results from broth micro-dilution while reducing the TAT to 3 h from several days which is required by the latter. Our study has demonstrated the micro-flow cell washing technique is a reliable, fast and compatible approach to replace centrifuge washing for sample pretreatment of Raman-AST and could be readily applied in clinical scenarios.

Sedimentation field-flow fractionation for rapid phenotypic antimicrobial susceptibility testing: a pilot study.

The increase in antibiotic resistance is a major public health issue. The development of rapid antimicrobial susceptibility testing (AST) methods is becoming a priority to ensure early and appropriate antibiotic therapy. To evaluate sedimentation field-flow fractionation (SdFFF) as a method for performing AST in less than 3 h. SdFFF is based on the detection of early biophysical changes in bacteria, using a chromatographic-type technology. One hundred clinical Escherichia coli strains were studied. A calibrated bacterial suspension was incubated for 2 h at 37°C in the absence (untreated) or presence (treated) of five antibiotics used at EUCAST breakpoint concentrations. Bacterial suspensions were then injected into the SdFFF machine. For each E. coli isolate, retention times and elution profiles of antibiotic-treated bacteria were compared with retention times and elution profiles of untreated bacteria. Algorithms comparing retention times and elution profiles were used to determine if the strain was susceptible or resistant. Performance evaluation was done according to CLSI and the ISO standard 20776-2:2021 with broth microdilution used as the reference method. AST results from SdFFF were obtained in less than 3 h. SdFFF showed high categorical agreement (99.8%), sensitivity (99.5%) and specificity (100.0%) with broth microdilution. Results for each antimicrobial were also in agreement with the ISO 20776-2 recommendations, with sensitivity and specificity of ≥95.0%. This study showed that SdFFF can be used as a rapid, accurate and reliable phenotypic AST method with a turnaround time of less than 3 h.

The Impact of Reporting the Same-Day Identification and Antibiotic Susceptibility Test Results on the Treatment of Bloodstream Infections.

The rise of antibiotic-resistant organisms necessitates the implementation of rapid identification (ID) and antibiotic susceptibility testing (AST) methods for patient management. We aimed to analyze how rapid ID and AST reporting influenced clinicians' treatment decisions. Bacteria were identified directly from positive blood cultures (BC) using serum separator tubes and MALDI-TOF MS. EUCAST rapid antibiotic susceptibility testing (RAST) method was performed for AST. The impact of rapid ID and AST reports on clinician treatment decisions was evaluated through clinical documentation. The appropriateness of antimicrobial therapy and interventions was assessed according to institutional antimicrobial prescribing guidelines, AST results, and clinical data. A total of 128 BC bottles from 86 patients underwent processing. The rapid ID method was successful in 105 (82.1%) bottles obtained from 76 patients. The rapid ID results were reviewed by the Infectious Diseases Team on the same day for 55 (72.4%) of the 76 patients. Following the evaluation, new treatments or interventions were recommended for 28 (36.8%) patients. RAST results were available for 24 patients. The susceptibility profile of seven patients was assessed by the Infectious Diseases Team on the same day. Antimicrobial treatment was escalated in four cases, and de-escalation was made in two based on RAST results. If all rapid results had been assessed, adjustments could have been made for eight (10.5%) and eleven (14.5%) more patients, according to ID and RAST results, respectively. Implementation of rapid ID and AST may contribute to patient management. Although rapid reporting was made, some results were not evaluated by the clinician on the same day, indicating that communication between the clinician and the laboratory needs to be strengthened.

Evaluation of the QMAC-dRAST System Version 2.5 for Rapid Antimicrobial Susceptibility Testing of Gram-Negative Bacteria From Positive Blood Culture Broth and Subcultured Colony Isolates.

Rapid antimicrobial susceptibility testing (AST) for bloodstream infections (BSIs) facilitates the optimization of antimicrobial therapy, preventing antimicrobial resistance and improving patient outcomes. QMAC-dRAST (QuantaMatrix Inc., Korea) is a rapid AST platform based on microfluidic chip technology that performs AST directly using positive blood culture broth (PBCB). This study evaluated the performance of QMAC-dRAST for Gram-negative bacteria using PBCB and subcultured colony isolates, comparing it with that of VITEK 2 (bioMérieux, France) using broth microdilution (BMD) as the reference method. We included 141 Gram-negative blood culture isolates from patients with BSI and 12 carbapenemase-producing clinical isolates of Enterobacterales spiked into blood culture bottles. QMAC-dRAST performance was evaluated using PBCB and colony isolates, whereas VITEK 2 and BMD were tested only on colony isolates. For PBCB, QMAC-dRAST achieved 92.1% categorical agreement (CA), 95.3% essential agreement (EA), with 1.8% very major errors (VMEs), 3.5% major errors (MEs), and 5.2% minor errors (mEs). With colony isolates, it exhibited 92.5% CA and 95.1% EA, with 2.0% VMEs, 3.2% MEs, and 4.8% mEs. VITEK 2 showed 94.1% CA and 96.0% EA, with 4.3% VMEs, 0.4% MEs, and 4.3% mEs. QMAC-dRAST yielded elevated error rates for specific antimicrobial agents, with high VMEs for carbapenems and aminoglycosides. The median time to result for QMAC-dRAST was 5.9 h for PBCB samples and 6.1 h for subcultured colony isolates. The QMAC-dRAST system demonstrated considerable strengths and comparable performance to the VITEK 2 system; however, challenges were discerned with specific antimicrobial agents, underlining a necessity for improvement.

Implementation of the EUCAST rapid antimicrobial susceptibility test (RAST) for carbapenemase/ESBL-producing Escherichia coli and Klebsiella pneumoniae isolates, and its effect on mortality.

With the rise in antimicrobial resistance, there is a growing demand for rapid antimicrobial susceptibility testing (RAST). In this study, we applied the EUCAST RAST method to ESBL/carbapenemase-producing Escherichia coli and Klebsiella pneumoniae isolates without using advanced identification systems and analysed the effect of this method on mortality rates Also the clinical impact of this method on patients infected with these bacteria and its effect on mortality rates were investigated. RAST was used for clinical blood cultures containing carbapenemase/ESBL-producing E. coli and K. pneumoniae without advanced identification systems (e.g. MALDI TOF), with preliminary identification by simple diagnostic tests (predicted RAST, or p-RAST), and its categorical agreement was investigated. The impact of the method on mortality was analysed by comparing the clinical data of patients whose blood cultures were subject to p-RAST (p-RAST group, n = 49) and those who were not subject to p-RAST (non-RAST group, n = 145). p-RAST results were analysed based on 539 antibiotic-bacteria combinations. Total error rates at 4, 6 and 8 h of incubation were 2.9%, 3.9% and 3.8%, respectively. In the p-RAST group, patients who did not receive appropriate antibiotics (29/45, 59.1%) were switched to appropriate treatment within 8 h at the latest. In contrast, in the non-RAST group, treatment of patients who received inappropriate antibiotics (79/145, 54.5%) could be changed after at least 24 h. Mortality rates were lower in the p-RAST group than in the non-RAST group (28.6% versus 51.7%, P = 0.005). p-RAST can be used safely in hospital laboratories with high rates of antimicrobial resistance and can reduce mortality rates by shortening the transition time to appropriate treatment.

Bacteriological profile and their antimicrobial susceptibility pattern among clinical suspected adult septicemia admitted patients: A study from tertiary care and teaching hospital

Objectives: The objective of this study is to determine the bacteriological profile and their antimicrobial susceptibility pattern among clinically suspected adult septicemia admitted patients and to study the correlation between the clinical presentation and inflammatory biomarkers (C-reactive protein [CRP] and Procalcitonin [PCT]) in the final outcomes of nosocomial sepsis in adult septicemia patients. Material and Methods: This is a descriptive cross-sectional study of 1-year duration from November 2022 to November 2023. The study has been approved from the Institutional Ethical Committee. Adult patients with a clinical suspected diagnosis of sepsis were admitted to the medicine ward of the tertiary hospital. Blood samples of adult sepsis suspected patients samples were taken for as a part of septicemia screening. Blood culture and sensitivity were performed according to standard guidelines. The sepsis inflammatory prognostic markers CRP and PCT test were evaluated according to manufactures instruction. Results: Out of 300 clinically suspected adult septicemia patients, 93 (31%) patients were isolated culture-positive sepsis while 207 (69%) patients were culture-negative sepsis. Gram-positive organisms were more (69%) as compared to Gram-negative organisms (31%). Staphylococcus aureus was predominant among Gram-positive organisms while Acinetobacter baumanii complex (9%) followed by Enterobacter spp. (8%) isolated in Gram-negative bacilli. Culture-positive sepsis patients have CRP level ≥3.2 mg/dL (80%) and the majority (80%) of these patients have reactive PCT. Conclusions: Successful treatment of bloodstream infection depends on early diagnosis and appropriate use of antimicrobial agents. Rapid identification results and antimicrobial susceptibility tests are essential for guiding clinicians in the selection of the most appropriate treatment for patients with bloodstream infections.

Comparison of Rapid Antibiotic Susceptibility Test Method Directly from Blood Culture Bottle with Standard Disc Diffusion Method

Aim: Early determination of antimicrobial susceptibility of sepsis pathogens is important. In this study, we aimed to compare the standard disc diffusion method with the rapid antimicrobial susceptibility testing (RAST) method performed directly from blood culture bottles. Material and Methods: Bacteria isolated from samples that gave a positive signal on the blood culture device between April 2019 and September 2019 were included in the study, and antimicrobial susceptibilities were determined by the standard disc diffusion method and the RAST method. Categorical agreement, small error, large error, very large error, and area of technical uncertainty ratios were recorded. Results: A total of 103 bacteria including 19 S. aureus, 10 Enterococcus spp. and 24 E. coli, 24 K. pneumoniae, 13 P. aeruginosa, and 13 A. baumannii were included in the study. When the RAST method was compared with the standard disc diffusion method, 100% agreement was found between the methods against imipenem, meropenem, gentamicin, and trimethoprim-sulfamethoxazole in E. coli isolates at all hours evaluated, and against meropenem in K. pneumoniae isolates at the 6th and 8th hour. For S. aureus and P. aeruginosa isolates, very major errors were found in the RAST results. For A. baumannii isolates, 100% agreement between methods was observed for many antibiotics. Conclusion: It was concluded that the RAST method is a simple and inexpensive test for life-threatening infections such as sepsis. It was also felt that similar studies should be carried out with a large number of isolates, as compliance rates vary depending on the bacteria tested.

Using the CLSI rAST breakpoints of Enterobacterales in positive blood cultures

ObjectivesThe objective of this study was to provide the clinic with rapid and accurate results of antimicrobial susceptibility testing for the treatment of patients with bloodstream infections. To achieve this, we applied the Clinical and Laboratory Standards Institute (CLSI) blood culture direct rapid antimicrobial susceptibility test (rAST) to assess the susceptibility of the most common Enterobacterales found in blood cultures. MethodsIn this study, we utilized the CLSI blood culture direct rapid antimicrobial susceptibility test to assess the susceptibility (rAST) of the most common Enterobacterales present in blood cultures. We chose this method for its simplicity in analysis, and our aim was to predict minimum inhibitory concentrations (MICs) using the rAST. As a benchmark, we assumed that Broth Macrodilution method (BMD) results were 100% accurate. For data evaluation, we employed the terms categorical agreement (CA), very major errors (VME), and major errors (ME). ResultsOur findings demonstrate that the CLSI rAST method is reliable for rapidly determining the in vitro susceptibility of Enterobacterales to common antimicrobial drugs in bloodstream infections. We achieved a concordance rate of 90% in classification within a 10-hour timeframe. We identified a total of 112 carbapenem-resistant Enterobacterales (CRE) strains, and there was no significant difference in the detection rate of CRE at 6, 10, and 16 hours. This suggests that CRE can be identified as early as 6 hours. ConclusionThe CLSI rAST is a valuable tool that can be utilized in clinical practice to quickly determine the susceptibility of Enterobacterales to antimicrobial drugs within 10 hours. This capability can greatly assist in the clinical management of patients with bloodstream infections.

Combining deep learning and droplet microfluidics for rapid and label-free antimicrobial susceptibility testing of colistin

Efficient tools for rapid antibiotic susceptibility testing (AST) are crucial for appropriate use of antibiotics, especially colistin, which is now often considered a last resort therapy with extremely drug resistant Gram-negative bacteria. Here, we developed a rapid, easy and miniaturized colistin susceptibility assay based on microfluidics, which allows for culture and high-throughput analysis of bacterial samples. Specifically, a simple microfluidic platform that can easily be operated was designed to encapsulate bacteria in nanoliter droplets and perform a fast and automated bacterial growth detection in 2 h, using standardized samples. Direct bright-field imaging of compartmentalized samples proved to be a faster and more accurate detection method as compared to fluorescence-based analysis. A deep learning powered approach was implemented for the sensitive detection of the growth of several strains in droplets. The DropDeepL AST method (Droplet and Deep learning-based method for AST) developed here allowed the determination of the colistin susceptibility profiles of 21 fast-growing Enterobacterales (E. coli and K. pneumoniae), including clinical isolates with different resistance mechanisms, showing 100 % categorical agreement with the reference broth microdilution (BMD) method performed simultaneously. Direct AST of bacteria in urine samples on chip also provided accurate results in 2 h, without the need of complex sample preparation procedures. This method can easily be implemented in clinical microbiology laboratories, and has the potential to be adapted to a variety of antibiotics, especially for last-line antibiotics to optimize treatment of patients infected with multi-drug resistant strains.

β-Galactosidase-triggered in situ synthesis of yellow emitting silicon nanoparticle and its application in visual detection of E. coli O157:H7 and drug susceptibility test

The sensitive detection of foodborne pathogenic and rapid antibiotic susceptibility testing (AST) is of great significance. This paper reports the enzyme-triggered in situ synthesis of yellow emitting silicon nanoparticles (SiNPs) and the detection of Escherichia coli (E. coli) O157:H7 in food samples and the rapid AST. The rapid counting of E. coli O157:H7 has been achieved through direct visual observation, equipment detection, and smartphone digitalization. A simple detection platform based on smartphone senses and cotton swabs has been established. Meanwhile, rapid AST based on enzyme-catalyzed SiNPs can intuitively obtain colorimetric samples. This paper established a system for bacterial enzyme-triggered in situ synthesis of SiNPs, with high responsiveness, luminescence ratio, and specificity. The detection limit for E. coli O157:H7 can reach 100 CFU/mL during 5 h, and the recovery efficiency ranges from 90.14% to 110.16%, which makes it a promising strategy for the rapid detection of E. coli O157:H7 and AST.

Accurate and rapid antibiotic susceptibility testing using a machine learning-assisted nanomotion technology platform.

Antimicrobial resistance (AMR) is a major public health threat, reducing treatment options for infected patients. AMR is promoted by a lack of access to rapid antibiotic susceptibility tests (ASTs). Accelerated ASTs can identify effective antibiotics for treatment in a timely and informed manner. We describe a rapid growth-independent phenotypic AST that uses a nanomotion technology platform to measure bacterial vibrations. Machine learning techniques are applied to analyze a large dataset encompassing 2762 individual nanomotion recordings from 1180 spiked positive blood culture samples covering 364 Escherichia coli and Klebsiella pneumoniae isolates exposed to cephalosporins and fluoroquinolones. The training performances of the different classification models achieve between 90.5 and 100% accuracy. Independent testing of the AST on 223 strains, including in clinical setting, correctly predict susceptibility and resistance with accuracies between 89.5% and 98.9%. The study shows the potential of this nanomotion platform for future bacterial phenotype delineation.

Persistent bacteremia caused by Ralstonia pickettii and Microbacterium: a case report

BackgroundRalstonia pickettii is a low virulent, gram-negative bacillus that is rarely associated with human infections and may cause bacteremia. Microbacterium species are gram-positive coryneforms that are generally considered as a contaminant in Gram staining of blood cultures, especially when the time to positivity is longer than 48 h. Both these bacterial species are emerging opportunistic pathogens that may occasionally cause serious infections and even life-threatening health conditions.Case presentationHere, we report the case of a patient with bacteremia caused by both R. pickettii and Microbacterium. We advocate for providers to order rapid antibiotic susceptibility testing, since our patient’s suffered two kinds of rare pathogens with the opposite of drug sensitivity results to imipenem.ConclusionsOur case present a patient suffered septic shock caused by R. pickettii and Microbacterium. Improving the antibiotic management based on the result of antimicrobial susceptibility tests is the key of successful treatment.

Rapid diagnostic of multidrug-resistant sepsis pathogens directly from blood culture bottles using MALDI-TOF and the EUCAST RAST

In this study, rapid diagnostic of multidrug-resistant (MDR) sepsis pathogens, directly from positive blood culture (BC) bottles, was evaluated by combining MALDI-TOF and the EUCAST Rapid Antimicrobial Susceptibility Testing (RAST). Carbapenemase production in Escherichia coli and Klebsiella pneumoniae isolates was also evaluated by RAST. From 171 positive BC bottles analyzed, 79 (46 %) MDR species, including E. coli (4/34, 12 %), K. pneumoniae (33/48, 69 %), Pseudomonas aeruginosa (12/12, 100 %), Acinetobacter baumannii (15/15, 100 %), and Staphylococcus aureus (14/37, 38 %) displaying resistance to beta-lactams, fluoroquinolones, aminoglycosides, and/or trimethoprim/sulphamethoxazole, were identified. In this regard, turnaround time of direct MALDI-TOF identification and RAST was < 7 h, which was significantly (p< 0.05) lower than our routine method. Carbapenemase detection by RAST displayed 100% sensitivity and 88.7 % specificity at 8 h. This protocol could offer advantages for the treatment and clinical outcomes of septic patients, improving the rapid diagnostic of sepsis by MDR pathogens.

A simplified version of rapid susceptibility testing of bacteria and yeasts using optical nanomotion detection.

We present a novel optical nanomotion-based rapid antibiotic and antifungal susceptibility test. The technique consisted of studying the effects of antibiotics or antifungals on the nanometric scale displacements of bacteria or yeasts to assess their sensitivity or resistance to drugs. The technique relies on a traditional optical microscope, a video camera, and custom-made image analysis software. It provides reliable results in a time frame of 2-4 h and can be applied to motile, non-motile, fast, and slowly growing microorganisms. Due to its extreme simplicity and low cost, the technique can be easily implemented in laboratories and medical centers in developing countries.