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Related Experiment Videos

Timely culture for mycobacteria which utilizes a microcolony method

D F Welch1, A P Guruswamy, S J Sides

  • 1Clinical Microbiology Laboratories, University of Oklahoma Health Science Center, Oklahoma City 73126.

Journal of Clinical Microbiology
|August 1, 1993
PubMed
Summary

A novel microscopic method for isolating mycobacteria significantly reduces detection time from an average of 23 days to 11 days. This microcolony examination allows for earlier presumptive identification of Mycobacterium tuberculosis and M. avium-M. intracellulare, improving clinical diagnosis.

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Area of Science:

  • Microbiology
  • Clinical Diagnostics
  • Mycobacteriology

Background:

  • Accurate and timely isolation of mycobacteria from clinical specimens is crucial for effective patient management.
  • Traditional methods for mycobacterial culture often involve lengthy incubation periods, delaying diagnosis and treatment initiation.

Purpose of the Study:

  • To evaluate a novel method for mycobacterial isolation using microscopic examination of microcolonies on Middlebrook 7H11 agar.
  • To compare the speed and accuracy of this microcolony method against conventional macroscopic examination techniques.

Main Methods:

  • Clinical specimens were cultured on thinly poured Middlebrook 7H11 agar plates (10x90 mm).
  • Plates were incubated and examined microscopically (x40-x180 magnification) for microcolony development.

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  • The microcolony method was compared to traditional culture methods using 270 mycobacterial isolates.

    • Main Results:

    • The microcolony method reduced the average time to mycobacterial detection to 11 days, compared to 23 days for the conventional method.
    • For Mycobacterium tuberculosis, the average time to positivity was 7 days with the microcolony method versus 17 days conventionally.
    • Presumptive identification of Mycobacterium tuberculosis and M. avium-M. intracellulare was achieved with high accuracy (83.5% and 85%, respectively) within 10-11 days.

    Conclusions:

    • Microscopic examination of microcolonies on Middlebrook 7H11 agar offers a faster alternative for mycobacterial isolation and presumptive identification.
    • Combining the microcolony method with conventional tube media can optimize recovery speed and sensitivity.
    • This method facilitates early detection of mixed mycobacterial infections and aids in selecting appropriate confirmatory tests.