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Free Content Rapid drug susceptibility testing of mycobacteria by culture on a highly porous ceramic support

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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.

Keywords: Anopore; rapid DST; tuberculosis

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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  • The International Journal of Tuberculosis and Lung Disease publishes articles on all aspects of lung health, including public health-related issues such as training programmes, cost-benefit analysis, legislation, epidemiology, intervention studies and health systems research. The IJTLD is dedicated to the continuing education of physicians and health personnel and the dissemination of information on tuberculosis and lung health world-wide.

    Certain IJTLD articles are selected for translation into French, Spanish, Chinese or Russian. They are available on the Union website

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