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Standardized disk-diffusion susceptibility test for Haemophilus influenzae.

J H Jorgensen, J C Lee

    American Journal of Clinical Pathology
    |March 1, 1977
    PubMed
    Summary
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    A new disk-diffusion method for Haemophilus influenzae susceptibility testing using supplemented Mueller-Hinton agar allows for routine clinical laboratory use. This practical approach accurately identifies ampicillin resistance in H. influenzae isolates.

    Area of Science:

    • Microbiology
    • Clinical Diagnostics
    • Antimicrobial Resistance

    Background:

    • Emergence of ampicillin-resistant Haemophilus influenzae necessitates improved susceptibility testing methods.
    • Standardized Kirby-Bauer disk-diffusion is challenging for fastidious organisms like H. influenzae.
    • Previous work established a simplified medium for quantitative dilution susceptibility testing.

    Purpose of the Study:

    • To evaluate a modified disk-diffusion method for routine susceptibility testing of Haemophilus isolates.
    • To assess the efficacy of ampicillin and chloramphenicol susceptibility testing using supplemented Mueller-Hinton agar.
    • To provide a practical method for hospital diagnostic laboratories.

    Main Methods:

    • Paired broth-dilution and disk-diffusion susceptibility tests were performed on 100 Haemophilus strains.

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  • Disk-diffusion tests utilized supplemented Mueller-Hinton agar incubated in an increased-CO2 atmosphere.
  • The method was adapted from the standardized Kirby-Bauer procedure.

    • Main Results:

    • Ampicillin-susceptible Haemophilus strains showed zone diameters ≥22 mm; resistant strains showed ≤18 mm.
    • All 100 strains were chloramphenicol-susceptible with zone diameters ≥30 mm.
    • The modified disk-diffusion method demonstrated clear differentiation of ampicillin resistance.

    Conclusions:

    • A modified disk-diffusion method using supplemented Mueller-Hinton agar is effective for routine susceptibility testing of H. influenzae.
    • This method facilitates accurate identification of ampicillin resistance in clinical isolates.
    • The technique is practical for hospital diagnostic laboratories and uses a clear medium.