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Development of a Broth Microdilution Method for Exebacase Susceptibility Testing.

Jun T Oh1, Jane E Ambler1, Cara Cassino1

  • 1ContraFect Corporation, Yonkers, New York, USA.

Antimicrobial Agents and Chemotherapy
|April 27, 2021
PubMed
Summary
This summary is machine-generated.

A new method using cation-adjusted Mueller-Hinton broth supplemented with horse serum and dithiothreitol (CAMHB-HSD) was developed for accurate exebacase minimum inhibitory concentration (MIC) determination. This validated method ensures reliable susceptibility testing for the novel antibacterial agent exebacase.

Keywords:
CF-301CLSIMICdirect lytic agentsendolysinexebacaselysin

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Area of Science:

  • Microbiology
  • Pharmacology
  • Clinical Diagnostics

Background:

  • Exebacase (CF-301) is a novel protein-based antibacterial agent, a lysin with antistaphylococcal activity, currently in Phase 3 clinical development.
  • Accurate and reproducible minimum inhibitory concentration (MIC) determination is crucial for clinical advancement.
  • Standard Clinical and Laboratory Standards Institute (CLSI) broth microdilution (BMD) methods showed limitations, including trailing endpoints and diminished activity with frozen panels.

Purpose of the Study:

  • To develop and validate an accurate and reproducible method for exebacase MIC determination.
  • To establish quality control (QC) ranges for exebacase susceptibility testing.
  • To gain CLSI approval for the modified testing method.

Main Methods:

  • A modified BMD method was developed using cation-adjusted Mueller-Hinton broth supplemented with 25% horse serum and 0.5 mM dl-dithiothreitol (CAMHB-HSD).
  • Preliminary quality control (QC) ranges were determined for Staphylococcus aureus ATCC 29213 and Enterococcus faecalis ATCC 29212.
  • A systematic study evaluated the impact of various parameters on susceptibility testing and assessed exebacase MIC distribution against clinical S. aureus isolates.

Main Results:

  • The modified CAMHB-HSD method resolved trailing MIC endpoints and maintained exebacase activity.
  • Preliminary QC ranges were established: 0.25 to 1 μg/ml for S. aureus ATCC 29213 and 16 to 64 μg/ml for E. faecalis ATCC 29212.
  • Data from systematic studies and QC testing supported the CLSI Subcommittee on Antimicrobial Susceptibility Testing (AST) approval of CAMHB-HSD.

Conclusions:

  • The modified BMD method using CAMHB-HSD provides accurate and reproducible exebacase susceptibility data.
  • This validated method is essential for the ongoing clinical development of exebacase.
  • Standardized testing and QC are vital for reliable antibacterial drug development.