Rapid drug susceptibility testing of mycobacteria by culture on a highly porous ceramic support
Abstract:BACKGROUND: Phenotypic, culture-based methods for drug susceptibility testing (DST) of Mycobacterium tuberculosis are relatively simple and may be particularly appropriate for resource-limited settings where tuberculosis (TB) is most prevalent. However, these methods can be slow and generate significant amounts of infectious waste. Low-cost digital imaging and a unique porous ceramic support for cell culture (Anopore) may offer opportunities to improve this situation.
OBJECTIVE: To test a rapid DST method based on fluorescence microscopy of mycobacteria grown for a few generations on Anopore.
DESIGN: Mycobacteria were cultured with and without drugs, and the resulting microcolonies were heat-killed and stained with the fluorogenic dye Syto16. Microscopy, image-capture with a charge-coupled device camera and digital processing were used to quantify the inhibition of growth by drugs. Rapid DST for rifampicin and isoniazid was performed for clinical isolates.
RESULTS: Mycobacteria could be cultured, killed, stained and imaged on Anopore. For DST, the Anopore method gave an accurate result in 3 days.
CONCLUSION: This is an unprecedented speed for culture-based DST for this group of organisms and results in minimal infectious waste (<20000 colony forming units). Analysis of mycobacteria by fluorescence and electron microscopy on Anopore also opens up research possibilities.
Document Type: Regular Paper
Affiliations: 1: Laboratory of Medical Microbiology, Twente Achterhoek, Enschede, The Netherlands; Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands 2: Laboratory of Medical Microbiology, Twente Achterhoek, Enschede, The Netherlands
Publication date: 2008-06-01
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