A novel machine-learning aided platform for rapid detection of urine ESBLs and carbapenemases: URECA-LAMP.

Journal: Journal of clinical microbiology
PMID:

Abstract

Pathogenic gram-negative bacteria frequently carry genes encoding extended-spectrum beta-lactamases (ESBL) and/or carbapenemases. Of great concern are carbapenem resistant , , and . Despite the need for rapid AMR diagnostics globally, current molecular detection methods often require expensive equipment and trained personnel. Here, we present a novel machine-learning-aided platform for the rapid detection of ESBLs and carbapenemases using Loop-mediated isothermal Amplification (LAMP). The platform consists of (i) an affordable device for sample lysis, LAMP amplification, and visual fluorometric detection; (ii) a LAMP screening panel to detect the most common ESBL and carbapenemase genes; and (iii) a smartphone application for automated interpretation of results. Validation studies on clinical isolates and urine samples demonstrated percent positive and negative agreements above 95% for all targets. Accuracy, precision, and recall values of the machine learning model deployed in the smartphone application were all above 92%. Providing a simplified workflow, minimal operation training, and results in less than an hour, this study demonstrated the platform's feasibility for near-patient testing in resource-limited settings.IMPORTANCEExtended-spectrum beta-lactamases (ESBL) and carbapenemases confer resistance to third-generation cephalosporins and carbapenems in pathogenic Gram-negative bacteria such as , , and . Conventional antimicrobial susceptibility testing is based on phenotypic methods, and results can take several days to be obtained. Current genotypic detection methods can be rapid but require expensive equipment and trained personnel. In this study, we present a novel machine learning-aided platform for the rapid detection of ESBLs and carbapenemases using Loop-mediated isothermal Amplification (LAMP). The validation of the platform demonstrated percent positive and negative agreements above 95% for all targets. The newly developed platform provided a simplified workflow, minimal technical training, and results in less than an hour. This study demonstrated the platform's feasibility for rapid testing of ESBL and carbapenemases in bacteria and urine specimens.

Authors

  • L Ricardo Castellanos
    Department of Pathology & Laboratory Medicine, Medicine, and Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada.
  • Ryan Chaffee
    Department of Pathology & Laboratory Medicine, Medicine, and Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada.
  • Hitendra Kumar
    Department of Mechanical and Manufacturing Engineering, University of Calgary, Calgary, Alberta, Canada.
  • Biniyam Kahsay Mezgebo
    Department of Pathology & Laboratory Medicine, Medicine, and Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada.
  • Pawulos Kassau
    Amhara Public Health Institute, Amhara Bahir Dar, Ethiopia.
  • Gisele Peirano
    Division of Microbiology, Alberta Precision Laboratories, Calgary, Alberta, Canada.
  • Johann D D Pitout
    Division of Microbiology, Alberta Precision Laboratories, Calgary, Alberta, Canada.
  • Keekyoung Kim
    Department of Mechanical and Manufacturing Engineering, Schulich School of Engineering, University of Calgary, Calgary, AB, T2N 1N4, Canada.
  • Dylan R Pillai
    Department of Pathology & Laboratory Medicine, Medicine, and Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada.