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Choi, Jungil,Yoo, Jungheon,Kim, Ki-jung,Kim, Eun-Geun,Park, Kyung Ock,Kim, Hyejin,Kim, Haeun,Jung, Hyunju,Kim, Taeyoung,Choi, Myungjin,Kim, Hee Chan,Ryoo, Sungweon,Jung, Yong-Gyun,Kwon, Sunghoon Springer-Verlag 2016 Applied microbiology and biotechnology Vol.100 No.5
<P>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.</P>
A rapid antimicrobial susceptibility test based on single-cell morphological analysis
Choi, Jungil,Yoo, Jungheon,Lee, Mincheol,Kim, Eun-Geun,Lee, Ji Soo,Lee, Seungok,Joo, Seik,Song, Sang Hoon,Kim, Eui-Chong,Lee, Jung Chan,Kim, Hee Chan,Jung, Yong-Gyun,Kwon, Sunghoon American Association for the Advancement of Scienc 2014 Science Translational Medicine Vol.6 No.267
<P>A rapid antibiotic susceptibility test (AST) is desperately needed in clinical settings for fast and appropriate antibiotic administration. Traditional ASTs, which rely on cell culture, are not suitable for urgent cases of bacterial infection and antibiotic resistance owing to their relatively long test times. We describe a novel AST called single-cell morphological analysis (SCMA) that can determine antimicrobial susceptibility by automatically analyzing and categorizing morphological changes in single bacterial cells under various antimicrobial conditions. The SCMA was tested with four Clinical and Laboratory Standards Institute standard bacterial strains and 189 clinical samples, including extended-spectrum β-lactamase–positive <I>Escherichia coli and Klebsiella pneumoniae</I>, imipenem-resistant <I>Pseudomonas aeruginosa</I>, methicillin-resistant <I>Staphylococcus aureus</I>, and vancomycin-resistant <I>Enterococci</I> from hospitals. The results were compared with the gold standard broth microdilution test. The SCMA results were obtained in less than 4 hours, with 91.5% categorical agreement and 6.51% minor, 2.56% major, and 1.49% very major discrepancies. Thus, SCMA provides rapid and accurate antimicrobial susceptibility data that satisfy the recommended performance of the U.S. Food and Drug Administration.</P>
( Kyung Ock Park ),( Haeun Kim ),( Hyejin Kim ),( Eun Hee Lee ),( Hyun Ju Jung ),( Jung Il Choi ),( Jungheon Yoo ),( Sung Hoon Kwon ),( Sung Weon Ryoo ),( Yong Gyun Jung ) 대한내과학회 2014 대한내과학회 추계학술발표논문집 Vol.2014 No.1
Background: Early diagnosis of TB is crucial for both clinically and epidemiologically. Sputum culture is the gold standard for diagnosis of Mycobacterium tuberculosis tuberculosis (TB). The MAC system immobilizes bacteria by using agarose in a microfi uidic culture chamber so that single cell growth can be tracked by microscopy. This study using DAC system for detection of M. tuberculosis isolates from sputum samples. Methods: 74 sputa samples (25 clinical sputa, 49 H37Rv spiking sputa) were included for the diagnosis of TB. All samples were decontaminated using the 4% N-acetyl-Lcysteine (NALC)-NaOH method. Results: In 25 clinical sputa samples, positive rate for Middlebrook 7H11 medium, Ogawa`s medium, and DAC system were determined as 1 (4.0%), 1 (4.0%), and 3 (12.0%), respectively. Negative rate for Middlebrook 7H11 medium, Ogawa`s medium, and DAC system were determined as 23 (92.0%), 23 (92.0%), and 21 (84.0%), respectively. And 1 (4.0%) contamination was observed in all culture methods. In 49 H37Rv spiking sputa samples, time to detection (TTD) was 5 to 13 days in MGIT 960 and 5 to 9 days in DAC system. In a total of 49 H37Rv spiking sputa samples, TTD according to cfu in Middlebrook 7H11 medium was 7 days (5-6 log10 cfu/ml) to 9 days (2-3 log10 cfu/ml) in DAC system. TTD according to cfu in Middlebrook 7H11 medium was 6 days (5-6 log10 cfu/ml) to 12 days (2-3 log10 cfu/ml) in MGIT 960. Conclusions: The DAC system detected the growth of M. tuberculosis more sensitivity than Middlebrook 7H11 and Ogawa`s media. Also, required time for true negative reports in less than 3 weeks. We concluded the DAC culture system is advantageous in terms of turnaround time.