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Rapid drug susceptibility test of Mycobacterium tuberculosis using microscopic time-lapse imaging in an agarose matrix

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Abstract

Tuberculosis (TB) is a major global health problem, and multi-drug-resistant TB (MDR-TB) and extensively drug-resistant TB (XDR-TB) are spreading throughout the world. However, conventional drug susceptibility test (DST) methods, which rely on the detection of the colony formation on a solid medium, require 1–2 months to the result. A rapid and accurate DST is necessary to identify patients with drug-resistant TB and treat them with appropriate drugs. Here, we used microscopic imaging of Mycobacterium tuberculosis (MTB) immobilized in an agarose matrix for a rapid DST. The agarose matrix, which was molded in a microfluidic chip, was inoculated with MTB, and TB drugs in liquid culture medium diffused throughout the agarose to reach the MTB immobilized in the agarose matrix. After the responses of MTB to drugs were tracked with an automated microscopic system, an image-processing program automatically determined the susceptibility and resistance of MTB to specific doses of TB drugs. The automatic DST system was able to assess the drug susceptibility of various drug-resistant clinical TB strains within 9 days with an accuracy comparable to that of conventional method. Our rapid DST method based on microscopic time-lapse imaging greatly reduces the time required for a DST and can be used to rapidly and accurately treat TB patients.

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Correspondence to Sungweon Ryoo, Yong-Gyun Jung or Sunghoon Kwon.

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This work was supported by the Korean Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea (grant numbers HI13C1468 and HI13C0866); the Institute for Basic Science (IBS) in Korea; the Pioneer Research Center Program through the NRF of Korea funded by the Ministry of Science, ICT & Future Planning (NRF-2012-0,009,555), an NRF grant funded by the Korean Government (2012M3A9B2030170); and the Nano Material Technology Development Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (2012M3A7A9671610).

Conflict of interest

J. Yoo, E.-G. Kim., K.O. Park, Y.-G. Jung, and S. Kwon at the submission of the paper were employed at QuantaMatrix Inc., which is commercializing the rapid DST technology. J. Yoo, E.-G. Kim., K.O. Park, Y.-G. Jung, and S. Kwon have equity interest in QuantaMatrix Inc.

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This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Jungil Choi, Jungheon Yoo, and Ki-jung Kim contributed equally to this work

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Choi, J., Yoo, J., Kim, Kj. et al. Rapid drug susceptibility test of Mycobacterium tuberculosis using microscopic time-lapse imaging in an agarose matrix. Appl Microbiol Biotechnol 100, 2355–2365 (2016). https://doi.org/10.1007/s00253-015-7210-0

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