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Related Concept Videos

Rapid Identification of Pathogens01:25

Rapid Identification of Pathogens

MALDI-TOF MS has transformed clinical microbiology by offering a rapid and reliable method for pathogen identification. The traditional approach to microbial identification typically involves time-consuming culture techniques and biochemical tests, which can delay the initiation of appropriate antimicrobial therapy. MALDI-TOF MS avoids these delays by using characteristic ribosomal protein mass patterns of microbial cells, enabling accurate species-level identification within minutes.Principle...
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Automated diagnostic analyzers have transformed clinical microbiology by providing rapid and reliable methods for pathogen identification and antibiotic susceptibility testing. Among these systems, the Vitek 2 is widely used because it automates the traditionally labor-intensive processes of microbial identification (ID) and antibiotic susceptibility testing (AST), delivering standardized and timely results that are essential for effective patient care.Microbial Identification with ID CardsThe...

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Rapid pathogen-specific phenotypic antibiotic susceptibility testing using digital LAMP quantification in clinical

Nathan G Schoepp1, Travis S Schlappi1, Matthew S Curtis1

  • 1Division of Chemistry and Chemical Engineering, California Institute of Technology, 1200 East California Boulevard, Pasadena, CA 91125, USA.

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|October 6, 2017
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Summary

Rapid antimicrobial susceptibility testing (AST) using digital nucleic acid quantification provides results in under 30 minutes. This ultrafast method aids in timely treatment decisions and combats antimicrobial resistance.

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Area of Science:

  • Microbiology
  • Molecular Diagnostics
  • Antimicrobial Resistance

Background:

  • Urgent need for rapid antimicrobial susceptibility testing (AST) to guide treatment and curb antibiotic resistance.
  • Current phenotypic AST methods are time-consuming, exceeding single patient visit durations.
  • Antimicrobial resistance is a growing global health threat due to antibiotic misuse.

Purpose of the Study:

  • To develop and validate a rapid phenotypic AST method for clinical samples.
  • To achieve AST results within 30 minutes for informed clinical decision-making.
  • To utilize digital nucleic acid quantification for precise measurement of bacterial response to antibiotics.

Main Methods:

  • Developed an ultrafast digital real-time loop-mediated isothermal amplification (dLAMP) assay.
  • Measured the phenotypic response of *Escherichia coli* in clinical urine samples to antibiotics.
  • Utilized SlipChip microfluidic devices for sample processing and analysis.
  • Compared dLAMP assay performance against a commercial digital polymerase chain reaction (dPCR) assay.

Main Results:

  • Achieved AST results for *E. coli* in clinical urine samples in under 30 minutes.
  • The dLAMP assay demonstrated high accuracy (AUC, 0.96) compared to dPCR (AUC, 0.98).
  • Successfully measured phenotypic antibiotic susceptibility directly from clinical samples.

Conclusions:

  • Rapid digital AST (dAST) using dLAMP is feasible for *E. coli* in urine samples.
  • This technology enables faster clinical decisions and improves antimicrobial stewardship.
  • Further development could extend dAST to other pathogens, antibiotics, and sample types.