scholarly journals 2136. Rapid Antimicrobial Susceptibility Testing using ATP Luminescence and Machine Learning Methods

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S723-S724
Author(s):  
Shunsuke Kawabe ◽  
Yuichi Uchiho ◽  
Hideyuki Noda ◽  
Atsushi Matsui ◽  
Hideki Niimi ◽  
...  

Abstract Background To prevent the spread of drug-resistant bacteria, a rapid and accurate antimicrobial susceptibility test (AST) is necessary. Recently, morphokinetic microscopy approaches have been reported as a rapid AST method. However, these still require several hours to obtain a minimum inhibitory concentration (MIC). Adenosine triphosphate (ATP) luminescence has also been reported as a rapid AST method that can detect bacterial growth more rapidly than morphokinetic approaches, since ATP in bacteria increases prior to bacterial division. In this study, we designed a new machine learning-based algorithm that predicts MIC rapidly, using a dataset that contains ATP luminescence patterns and conventional MICs determined by turbidity. Essential agreement (EA) rates between rapid and conventional MIC were then evaluated. Methods Sixty-three strains of E. coli (ATCC 25922 and clinical isolates from Toyama University Hospital) were tested. Bacterial suspensions were diluted 500-fold in Mueller–Hinton broth from 0.5 McF solutions, and the final concentration of bacteria was 3×105 CFU/mL. The suspensions were dispensed into a 96-well microplate, which had 12 antimicrobials in two-fold dilution series, and the microplate was incubated at 35°C. At each measurement time point, the amount of ATP in a 10 μL aliquot from each well was evaluated by our original measurement system, which can sensitively detect ATP luminescence equivalent to a single bacterium. After 22 hours, MIC was determined conventionally by measuring turbidity. A rapid MIC for each bacterium was estimated by the algorithm based on the dataset consisting of the rest of the 62 strains (leave-one-out cross validation). Results Table 1 shows the EA rate for the 12 antimicrobials; EA rates > 90% were achieved for 7 antimicrobials in 2 hours and for 12 antimicrobials in 3 hours. In 6 hours, an average EA rate > 97% was achieved. Conclusion Using the dataset, our new machine learning-based algorithm predicted MIC rapidly within 2 hours with an EA rate > 90% for 7 antimicrobials. The rapid AST detected by the ATP luminescence method will contribute toward both appropriate antimicrobial treatment and reduction in medication and admission charges. In the future, other species of bacteria will be evaluated by our ATP method. Disclosures All authors: No reported disclosures.

2020 ◽  
Vol 30 (Supplement_5) ◽  
Author(s):  
L Tilouche ◽  
N Haddad ◽  
S Boujaafar ◽  
R Elaissi ◽  
S Kahloun ◽  
...  

Abstract Background The discovery of antibiotics revolutionized medicine in the 20th century, however the emergence of extensively drug-resistant bacteria constitute a growing problem in our hospitals in Tunisia and across the world. This study aims to evaluate a screening program for Carbapenemase Producing Enterobacteriaceae (CPE) and vancomycin-resistant enterococci (VRE) undertaken by the Microbiology Laboratory of Sahloul University Hospital. Methods A descrptive and retrospective study was carried out between 01st April 2018 and 31th December 2019 in the university hospital Sahloul, in East Coast of Tunisia. The screening was based on rectal swabs; it concerned Medical Intensive Care Units, the postoperative unit, the paediatric and the nephrology departments. The rectal swabs were seeded on the chromID CARBASMART and chromID VRE agars(biomerieux, France)in search of CPE and VRE respectively. The identification and the antibiotic susceptibilities testing were performed using the Vitek2 System (biomerieux,France) Results In total, 191 patients were screened. Among them, 56% were admitted in Intensive Care Units, 9.95% in postoperative unit, 30.9% in paediatric department and 3.14% in the nephrology department. A total of 38 extensively drug-resistant bacteria were isolated: 20 CPE and 18 VRE. Among isolated CPE, 17 were identified as Klebsiella pneumonia: The ConfirmationKPC/MBLkit (RoscoDiagnostica, Denmark) supplemented with a disc of Temocillin showed that 7 strains produced metallo-carbapenemase and 10 strains produced OXA 48 carbapenemase. The other isoloted CPE were OXA-48 producers. All VRE were identified as Enterococcus faecium, Their Vancomycin and Teicoplanine MICs were greater than 32 mg/L. Conclusions Mastering the spread of extensively drug-resistant bacteria involves a multidisciplinary preventive strategy. It must include strict application of hygiene measures, early detection and isolation of carriers and rationalization of antibiotic use. Key messages the emergence of extensively drug-resistant bacteria constitute a growing problem that`s why Carbapenemase Producing Enterobacteriaceae and vancomycin-resistant enterococci screening is crucial. Anti microbial agents use must be rationalized.


Author(s):  
Hamza A. J. ◽  

Coagulase-negative staphylococci (CoNS) were formally believed to be non-pathogenic but recent studies have implicated them as agents of some diseases. This study was designed to investigate the occurrence and antimicrobial susceptibility profile of CoNS isolated from local yoghurt hawked in Gombe metropolis, Nigerian. A total of 177 local yoghurt samples from Gombe metropolis were investigated for the presence of CoNS using standard microbiological methods and confirm using Microgen Staph ID kit. The antibiotics susceptibility profile of the confirmed isolates was determined using disc diffusion method. Out of the 177 yoghurt samples, a total 24 (13.56%) were identified as CoNS which include: S. chromogenes 11(45.83%), S. haemolyticus 5(20.83), S. hominis 3(12.50%), S. warneri 3(12.50%), S. cohnis 1(4.17%) and S. ciuri 1(4.17%). Antimicrobial susceptibility testing revealed that the isolates were highly resistant to ampicilin (75%) and cefoxitin (54.2%) but very sensitive to gentamicin (100%), vancomycin (95%) and chloramphenicol (100%). The study reveals the presence of CoNS in the study area that are highly resistant to conventional antibiotics. Thus, to ensure public health safety and to avoid transmission of drug resistant bacteria, there is the need to educate yoghurt producers, vendors and consumers on the importance of food safety by the relevant authorities. Keywords: Coagulase negative staphylococci, Local yoghurt, Antimicrobial Resistance


2021 ◽  
Vol 15 (05) ◽  
pp. 665-671
Author(s):  
Ozge Unlu ◽  
Berken Rabun Ersoz ◽  
Ayse Istanbullu Tosun ◽  
Mehmet Demirci

Introduction: Klebsiella pneumoniae sequence type 258 (ST258) strains are globally distributed multi-drug resistant pathogens and can spread rapidly throughout the world, causing severe healthcare-associated invasive infections with limited antimicrobial treatment options. The aim of this study was to reveal the incidence of Klebsiella pneumoniae ST258 strains among the intensive care unit patients in a university hospital in Istanbul. Methodology: Consecutive nonreplicated 83 K. pneumoniae strains were isolated from various clinical samples of intensive care unit patients admitted to a university hospital in Istanbul, between November 2016 to December 2018. Bacterial identifications were performed via VITEK2. Antimicrobial susceptibility tests were conducted with Kirby Bauer’s disc diffusion test except for colistin which was performed with broth microdilution. Real-time PCR method was utilized in order to reveal ST258 positivity among the strains. Results: Antimicrobial susceptibility results revealed that 56 (67%) K. pneumoniae strains were carbapenem-resistant. Real-time PCR results demonstrated that 15 out of 83 (18%) K.pneumoniae strain were ST258. According to antimicrobial susceptibility test results of ST258 strains, 8 were found as carbapenem-resistant whereas 7 were found as carbapenem susceptible. 3 out of 8 (37.5%) carbapenem-resistant ST258 strains were found as resistant against all antibiotics tested. Conclusions: Our study revealed that K. pneumoniae ST258 which caused severe infections worldwide so far has also spread to Istanbul. We believe that rapid molecular methods for monitorization of these clones are useful. our results showed that ST258 is not linked to a multi-resistant strain and suggested that it does not contribute to multi-resistance formation alone.


2018 ◽  
Vol 2018 ◽  
pp. 1-7 ◽  
Author(s):  
Moti Tolera ◽  
Degu Abate ◽  
Merga Dheresa ◽  
Dadi Marami

Nosocomial infections remain a major cause of mortality and morbidity worldwide. Despite the highly specialized interventions and policies, the rate of infection is still high due to the emergence of antimicrobial-resistant bacteria. This study described the prevalence of bacterial nosocomial infections and antimicrobial susceptibility pattern of isolates among patients admitted at Hiwot Fana Specialized University Hospital, Eastern Ethiopia. A hospital-based cross-sectional study was conducted among 394 nosocomial infection-suspected patients from March 2017 to July 2017. Data were collected using a structured questionnaire. Specimens from the respective site of infections were collected and examined for the presence of pathogenic bacteria and their antimicrobial susceptibility using standard culture and serological tests. Data were summarized using descriptive statistics. The prevalence of culture-confirmed bacterial nosocomial infection was 6.9% (95% CI: 4.3–7.9). Staphylococcus aureus (18.5%) was the most common isolate followed by Escherichia coli (16.7%). S. aureus showed 80% resistance to chloramphenicol and erythromycin, and 70% to cephalexin and tetracycline, respectively. A methicillin-resistant S. aureus made up 88.9% of all S. aureus isolates. Pseudomonas aeruginosa showed 83.7% resistance to each of ceftazidime and cephalexin, and 66.7% to chloramphenicol. The most common multidrug-resistant isolates were P. aeruginosa (30.4%) and S. aureus (21.7%). The prevalence of nosocomial infections in this study was comparable with other findings; however, the high rates of antimicrobial resistant isolates represent a substantial threat to the patients, communities, health care providers, and modern medical practices. Bacterial nosocomial infection treatment should be supported by culture isolation and antimicrobial susceptibility testing.


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